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Certain HIV positive, but negative tests (Page 61)

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January 28th, 2013
Experienced User
long time havent been here.
just an update
the hyperthermia did nothing for me except my symptoms has been worsen.

I really think it is not HTLV
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replied January 29th, 2013
Extremely eHealthy
Hope,

I've been thinking about you often, sorry to hear that the hyperthermia didn't work for you. Can you tell us about your experience with hyperthermia please? And also which of your symptoms worsened?

Best wishes.
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replied January 29th, 2013
Experienced User
Ustas, scared49 , Drank.. ??? Where are you guys how are you guys doin? Any progress? Please respond or give us an update.

My symtoms are still here..

Thing in throat like something is stuck in theere.. bone, hair,
Follucollitis and icthy skin. Red spots then scab. Mostly on chest and arms and face.
Muscle pain legs always sore and other body muscles sore also
Joint pain mostly hips and legs
Sensitive neck. Can get sore easily
Stomach cramps . Irregular bowel movents



My gf is expieriencing a lot of the same symtoms..
Shes going to a lot of dr appts. And hasnt had her period in about 4 motnhs and not preggo. She has irregular hormones. And inlarged glands or something in her head. Dr said its something like a small tumur! Im so scared..

I really wanna test for mycoplasma and other bacterial infections. I feel like this is getting worse
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replied January 29th, 2013
Experienced User
Next month on feb 19th marks a year since my exposure
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replied January 29th, 2013
Extremely eHealthy
D-man,

Sorry that you are still suffering - we all are. I'm sure that the people we haven't heard from aren't doing any better. The doctors don't want to accept that this is a new disease that is making people miserably sick. And if it progresses like HTLV, we could end up dead from this, and the medical authorities wouldn't care to even count us as a statistic - our death certificate would say something vague like "death from sepsis", as was done when Doctor Bob died. I have all of your symptoms too.

Best wishes.
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replied January 29th, 2013
Experienced User
D-Man,Tony,Usts, scared49, onechanchance How are you all.
D-Man as Tony said i too have all your symptoms of legweakness,Muscle pain legs always sore and other body muscles sore also
Joint pain mostly hips and legs
Sensitive neck. Can get sore easily
Stomach cramps . Irregular bowel movents.
Blurred vision. Also of late i get feel andcrinkling
noise at the back of my neck. feels like something happening with my CSF fluide.I still suspect HTLV but docs, HTLV reaserch forums dont believe HTLV is the cause.
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replied January 29th, 2013
Extremely eHealthy
AdviseMe,

Good to hear from you, man.

1. We've all tested (negative) for HTLV, but there's no formal window specified for HTLV (like the three month window for HIV).

2. We all have symptoms of HTLV.

3. Papers exist stating that patients in South America are showing up with HTLV symptoms but still test negative on multiple testing.

There's not a doctor in America (even who specialize in HTLV) who is going to admit this is HTLV. Obviously whether this is HTLV or not, we are dealing with a new epidemic that no medical person wants to acknowledge or deal with. Like HIV in the early 80s, not until people are filling the hospitals or dropping dead in large numbers will the medical authorities become concerned.

Best wishes.
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replied January 29th, 2013
Experienced User
Please check this and tell me what u think.

Researchers from the Blood Center of Ribeiro Preto, connected to the Hospital of the Faculty of Medicine of Ribeiro Preto (HCFMRP) USP, in partnership with Gene ID SA have developed a molecular diagnostic kit to confirm infection by the retrovirus HTLV 1 and 2 in case of positive results of tests done on the routine serologic blood centers and blood banks. Confirmatory testing domestic, and more sensitive, which allows to reduce the window period (the time between infection and antibody production against retroviruses), will cost 17 times less than that currently used, called Western blot (WB) which is imported.
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replied January 29th, 2013
Experienced User
there is this new serology test in brazil for htlv. go to youtube search watch?v=Kn6AqMXVeDY
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replied January 30th, 2013
Extremely eHealthy
AdviseMe,

Interesting news, thanks! I found it interesting because the physicians in the Brazilian video are Japanese, where HTLV is endemic.

Best wishes.
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replied January 29th, 2013
Experienced User
Tony,

Hey so I had a weird situation this weekend and wanted to see if anyone else had a similar situation. My symptoms while still present were much better. But after a few glasses of wine the night before, symptoms got much worse... Muscle , joint, chest pains, tiredness, etc

I guess It makes sense... Immune system weakened from alcohol and symptoms increased, maybe the inverse will be true. Rest... Etc.

I don't know ... Just an interesting dynamic... Anyone know of diseases that respond that quickly to stress/ alcohol?
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replied January 30th, 2013
Extremely eHealthy
W,

Myself and everyone I infected no longer have any tolerance for alcohol. I'm sure that's yet another symptom we all share. Having a drink now would basically send me to bed.

Best wishes.
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replied January 29th, 2013
Experienced User
Hey guys, sorry for not responding In a while. I've been reading the forum and sorry you all haven't found anything out yet. I'm too still searching and have just been severely depressed since all this happened. It's late now and I' have to work in the morning I'll check back tomorrow and let you guys know what all I've done and where I'm at. Got a lot of catching up to do. Again I'm sorry for not responding in a while, just been severely depressed. I'll talk to you guys tomorrow goodnight everyone and god bless you all.
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replied January 30th, 2013
Experienced User
Wyoming, stress and alcohol is probably bad for everyone with some kind of a disease.

The days after drinking alcohol I am extremely weak.

Have you guys tested for allergies?
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replied January 30th, 2013
Experienced User
e Office of Institutional Communication of the Blood Center of Ribeiro Preto

Researchers from the Blood Center of Ribeiro Preto, connected to the Hospital of the Faculty of Medicine of Ribeiro Preto (HCFMRP) USP, in partnership with Gene ID SA have developed a molecular diagnostic kit to confirm infection by the retrovirus HTLV 1 and 2 in case of positive results of tests done on the routine serologic blood centers and blood banks. Confirmatory testing domestic, and more sensitive, which allows to reduce the window period (the time between infection and antibody production against retroviruses), will cost 17 times less than that currently used, called Western blot (WB) which is imported.

An estimated 2.5 million people in Brazil have the retroviruses but only between 1% and 5% develop some kind of disease. The price of imported WB test is about $ 170.00 and also, in many cases, can not solve the patient diagnosis. Due to price sensitivity and the fact it is not mandatory, only a few institutions perform the confirmatory test. The national under development, uses a different methodology and beyond more efficient, will cost about $ 10.00, ie, only 6% of the value of imports. Another factor as important as the cost is to reduce the window period, which now ranges from 36 to 72 days.

Since 1993, blood centers are tests to identify blood donors among possible carriers of retroviruses HTLV. If the first test for the presence of HTLV constada, the stock is discarded and the donor is called by the medical staff of blood banks. But some institutions make to reduce doubts about the initial result, another test to confirm the result of the first. But most institutions does not perform the second test, considering only the result of the first.

"These factors allow the safe completion of diagnosis for proper counseling and treatment of patients, increased security in transplants and blood transfusions and prevent vertical transmission (mother to child), the main route of transmission," explains the researcher Mauricio Rocha, Laboratory of Molecular Biology of the Blood Center, led by researcher Simone Haddad. The coordination of the research is Professor Dimas Tadeu Covas, of the Faculty of Medicine of Ribeiro Preto (FMRP) USP.

Infected
Studies indicate that in the world there are about 20 million people infected with HTLV. Of these, approximately two and a half million are in Brazil, which has the largest absolute number of infected worldwide. Between 1% and 5% of this total will develop some disease related to HTLV. Among them, myelopathy tropical spastic paraparesis associated with HTLV, one prevalent in Brazil, for which there is no cure, but if discovered early-stage treatment provides an improvement in the patient's condition.

"It is a neurological disorder that affects the bone marrow in adults between 50 and 60 years, and that leads to muscle weakness of the legs. The person has difficulty walking and difficulty in controlling urine. It is a slowly progressive disease. It is a disease with rapid evolution, the patient takes many years to need wheel chair. The infection is transmitted by several mechanisms, including, breast milk, blood transfusion and sexual intercourse. There is no specific treatment. What you do is physical therapy and other measures for rehabilitation, "explains neurologist HCFMRP, Osvaldo Massaiti Takayanagui expert in treating the disease.

The next step in the research is to test blood samples from Salvador (Bahia), South Africa and Peru. "The proposal is to expand the number of exams from other regions to ensure the efficiency of the test. The samples tested indicate 100% efficiency, however, other samples need to be examined, because no diagnostic test is 100% efficient, "says Rocha. In this first phase were conducted 180 tests on samples from the database of the Blood Center of Ribeiro Preto.

The Financier of Studies and Projects (FINEP) is investing 1m in the production of national confirmatory test. The company Gene ID will be responsible for producing the kit, on completion of the survey. "With lower price and more efficient outcome, we hope that after completing all steps of the research kit to be used in routine diagnostic confirmatory of all blood centers in the country," said Rocha.

More information (16) 2101-9350, the Office of Institutional Communication of the Blood Center of Ribeiro Preto
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replied January 30th, 2013
Extremely eHealthy
AdviseMe,

Thanks for the excellent article! Not only because there's a serious need in Brazil for a cheaper, more reliable, shorter window test, but also because it mentions the window period being 36 to 72 days (recall blood banks use 51 days as their window period). I can't tell you how many of my friends traveled to Brazil to get laid, and now I learn that it has the HIGHEST number of HTLV infections in the world !!!

Best wishes.
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replied January 30th, 2013
Experienced User
SANTA ANA, Calif. - Monday, January 28th 2013 [ME NewsWire]

(BUSINESS WIRE)- MP Biomedicals, LLC announced today that it has submitted a BLA for FDA approval of the MP Diagnostics HTLV Blot 2.4, a Western blot for confirmatory testing and viral typing of blood donors and patients.

Additionally, MP Biomedicals plans to initiate an open IND study in February, which will allow for investigational use (IUO) of the assay on donor and patient samples while the product is under FDA evaluation. The American Red Cross is among the study participants. U.S. labs interested in participating may contact Karen Luthart for more information (see above).

“The HTLV Blot 2.4 represents a significant milestone for our company and it will be the first licensed confirmatory test for donor notification and counseling of HTLV infections in the U.S.,” said Milan Panic, Founder and Chief Executive Officer of MP Biomedicals. The assay is currently CE Marked and registered as a diagnostic device in 10 countries around the world.

An important aspect of the HTLV Blot 2.4 is the incorporation of unique recombinant proteins from the envelopes of both HTLV-I and HTLV-II viruses. This feature greatly enhances sensitivity and specificity, providing clinical and donor labs with reliable results for both confirmation and differentiation between HTLV-I and HTLV-II infections. This information is necessary when counseling infected donors, because of the different health implications of the two HTLV viral types.

HTLV is a retrovirus which infects predominantly T-lymphocytes in the human body. HTLV Type I is endemic in the Caribbean, Japan, South America, and parts of Africa. HTLV Type II is found among Native Americans. Although the vast majority of infected individuals are asymptomatic carriers, HTLV infection is known to cause Adult T-cell leukemia/lymphoma (ATLL), and HTLV-I-associated myelopathy (HAM) in some patients. HTLV can be transmitted through unprotected sexual contact, from an infected mother to her baby by prolonged breast feeding, sharing of needles by drug users, and transfusion of infected donor blood.
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replied January 30th, 2013
Extremely eHealthy
This is the second company in ten months to get approval for HTLV testing (Avioq, Inc did so last March). If HTLV is so rare, why are all these companies jumping into the market?

Best wishes.
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replied January 30th, 2013
Experienced User
You welcome tony. With all these test coming i hope some day they will find some treatment too or possible cure to this horrible disease.
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replied January 30th, 2013
Experienced User
You welcome tony. With all these test coming i hope some day they will find some treatment too or possible cure to this horrible disease.
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replied January 30th, 2013
Extremely eHealthy
AdviseMe,

There's definitely a sequence of events that needs to happen:

1. There needs to be a conclusive, reliable, known window HTLV test.

2. Everyone needs to be routinely tested for HTLV, just as is being done for HIV.

3. Once the medical authorities realize how many people are infected with HTLV, hopefully the ball will start rolling to treat it.

If you consider how long it took for each of these steps to occur for HIV, we have a long road ahead of us, and quite frankly, if people like Alberto can develop leukemia within 7 years, we are in big trouble.

I had forgotten how HIV devastated the hemophiliac populations until I read an article "Defective human T-cell leukaemia virus type (HTLV) genomes: no evidence in serologically indeterminate german blood donors but new type detected in established cell lines". The article mentions that hemophiliacs are also affected by HTLV.

Best wishes.
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replied January 30th, 2013
Experienced User
Hello everyone, well it's been a battle this past year and 2 months and counting and still no diagnosis. I strongly encourage EVERYONE on this forum to consider looking into gastrointestinal disorders Chrons Disease, Diverticulitis, IBS, Ulcerative Colitis, IBD, Liver, Gallbladder, and Pancreas Disorders. This is what I've been doing the past year now. I've had 4 Ultasounds of my abdomen, they've shown nothing. 10 yes 10 CT Scans of my abdomen and pelvis with and without contrast some have shown slightly enlarged Mesenteric Lymphnodes approx. 10 mm. One back in September showed a right heptic leison 3mm it said it was to small to characterize statiscally most likely a cyst. I just had another one done on jan. 21st and it said the liver leison is now 4mm. I told the doctor my concerns and he's basically blowing it off right now. I'm concerned this is some form of cancer just because of of my symptoms but its not showing on the ct scans. I had a MRI of my spine done back in August and it didn't show anything either. I've seen 2 different Gastroenterologists, Urologist, ENT Dr., Orthropedic, Pcp, and I've got nowhere. I forgot to mention I did have my Gallbladder removed because one of the drs thought that was my problem it wasn't. My next thing now is a Video Pill Endoscopy on Feb. 6 th because my Gastro dr. Now suspects Chrons Disease so we'll see. After that he's sending me to get my Pancreas evaluated. I'm so angry and depressed right now these drs are a waste all they care about is collecting money from the insurance companies. There's not a doubt this woman gave me something but I'm wondering if maybe there was something developing that I've been ignoring and just blaming it on this exposure. Maybe all of us I don't know. Anyways here's a list of my current symptoms...

Abdominal pains and discomfort
Irregular bowel movements
Light colored stools
Different colored stools
Moucus covered stools
Discomfort under right rib cage (4mm leison???)
Itchy skin especially at night, upper chest collarbone area
Sore knees
Sore legs
Muscle weakness
Fatigue
Insomnia
Muscle waisting in forearms
Joint pain
Dry mouth
Sticky saliva
Lines in fingernails
Middle back pain
Low back pin
Dark eyelids
Leg Weakness
Muscle tenderness
Lump feeling in throat
Repeated sinus infections
Sore throat off and on
Depression

I'll keep you all posted on whatever test results I have, good luck everyone.
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replied January 30th, 2013
Extremely eHealthy
Scared,

I appreciate all of the doctors' visits and testing that you have had done. We both got exposed to something around the same time (15 months for me) and I have all of your symptoms, including liver pain, fingernail lines, forearm wasting (and everywhere else), dark eyelids, etc. I feel like I have AIDS without having HIV, that's the best way to describe it, since there's SO MANY SYMPTOMS that I just feel like I am falling apart and dying everyday.

The amount of testing you've had is amazing, and I'm still sticking to my guns that we have HTLV. Can you please have all three tests for HTLV done (antibody, PCR, and Western Blot / Line Immunoassay) ???

Best wishes & good luck as well.
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replied January 30th, 2013
Extremely eHealthy
Scared,

Please give me a call tonight - I will leave my phone number in a private message to you.

Best wishes.
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replied January 30th, 2013
Experienced User
Hey Tony, it's funny you said that I feel like I have aids as well, but after 15 negative tests I let that go. I did have a negative HTLV antibody test I forgot to mention that, multiple I mean mutiple labs done WBC was low a couple of times. I to feel like I'm dying and I'm trying to stay strong for my family but man this really sucks. I don't even want to be at work or around people anymore.
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replied January 30th, 2013
Experienced User
Tony great idea, guys if any of you want to talk about anything I'm all ears. Here's my phone # (386)-748-6675. I try to get to sleep by about 10 or 11 p.m. E.T. You guys are welcome to call me if you want.
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replied February 1st, 2013
Extremely eHealthy
It was great talking to you on the phone, thanks! I'm glad that we have the same thoughts about what's going on with our health and the idiocy of doctors think that we are insane.

Best wishes.
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replied February 3rd, 2013
Experienced User
Hey man, anytime I just wish we could all find a dr. that takes us seriously. It's really sad that our medical field is the way it is really sad. I wish the best for everyone on this forum I really do its in gods hands guys he'll lead the way just have faith. I'm going to try and enjoy the superbowl tomorrow since I'm a die hard 49er fan. Good luck to everyone and ill post any test results that I have.

Good luck friends,
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replied February 3rd, 2013
Experienced User
htlv serointermediate
Scared49, tony, D-man and everyone in this forum. we all been trying in our own way as to find what is causing us all our symptoms.

I came through this paper from ncbi on HTLV serointermediate.it also explains enefficencies of other HTLV tests.sorry for this long post.

The first detection of the Human T-lymphotropic virus type I (HTLV-I) in 1980 marked the identification of the first human retrovirus [1]. While approximately 95% of HTLV-I infected individuals will remain asymptomatic for their lifetimes, roughly 5% of these patients will develop disease. Numerous diseases have been associated with the virus, including adult T-cell leukemia (ATL), HTLV-associated myelopathy/tropical spastic paraparesis (HAM/TSP), HTLV-I uveitis, and HTLV-I-associated infective dermatitis. In Japan, an estimated 1.2 million HTLV-I carriers have been identified, and more than 700 cases of adult T-cell leukemia (ATL) are diagnosed each year [2,3]. Since the initial detection and isolation of HTLV-I, HTLV research has expanded and led to the development of more sensitive and specific methods of anti-HTLV-I antibody detection. In 1984, a cell membrane immunofluorescence assay was used to detect antibodies to the HTLV-I surface protein Env. A few years later, indirect immunofluorescent assays for HTLV p19 were developed [4,5]. In 1988, radioimmunoprecipitation assays (RIPA) were used to detect these antibodies in patients who had not tested positive by other immunofluorescent methods [6]. Eventually, enzyme-linked immunosorbent assays (ELISA) became commonplace in anti-HTLV-I antibody detection, as well as the use of Western Blots (WB) to confirm infectivity [7,8]. More recently, we have developed a luciferase immunoprecipitation systems (LIPS) assay that has increased specificity and sensitivity in anti-HTLV-I antibody detection [9]. Collectively, these new methodologies have helped to shed light on the prevalence of the virus, indicating that approximately 1520 million individuals worldwide are infected with HTLV-I. Specifically, the virus is known to be endemic in Japan, several Caribbean countries, sub-Saharan Africa, South America, and areas of Iran and Melanesia.

Among diseases associated with HTLV-I is a chronic, progressive, neurodegenerative disease termed HAM/TSP. Since the initial report of the association between HTLV-I and HAM/TSP, more associations were discovered between HTLV-I and diseases including HTLV-I uveitis, bronchopneumonitis, arthritis, polymyositis, and various other inflammatory conditions [3,10,11]. As correlations of HTLV-I with a range of medical conditions were being demonstrated, information regarding the transmission of the virus was reported as well. HTLV-I infection requires cell-to-cell contact with infected T cells. Transmission may occur via sexual contact, vertical transmission (i.e., breast milk), and transfusion with infected blood [1012]. Specifically, there is an increased risk for developing HAM/TSP associated with transfusion. In Japan, 20% of HAM/TSP patients reported a history of blood transfusion as opposed to 3% of normal donors (p < 0.001) [11].

Due to this increased risk of disease with blood transfusion, many screening methods have been employed to detect HTLV-I infected donors. Blood bank screening for the virus was implemented in Japan in 1986, the United States in 1988, France in 1991, and the Netherlands in 1993 [10,11]. These screenings were initiated once proof of increased risk for developing HAM/TSP was definitively associated with transfusion. Numerous procedures exist for anti-HTLV-I antibody detection, including ELISA, RIPA, and particle agglutination. Standard screening assays used by blood banks in the United States include initial testing by an enzyme immunoassay (EIA), followed by confirmation of seropositivity by WB [12].

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2.Defining and Identifying Seroindeterminates
The WB used to confirm seropositivity in an EIA positive patient sample utilizes recombinant proteins specific for HTLV-I/II Env glycoproteins incorporated into the WB strips. These recombinant proteins increase the sensitivity of the blot, as well as differentiate between HTLV-I and HTLV-II. The specific sensitivity of an EIA assay is approximately 95% and the EIA specificity averages 98% while the sensitivity of the WB is 97.1% with a specificity of 97.5% [13,14]. A sample is classified as HTLV-I positive if it meets the established criteria (see Figure 1). According to the World Health Organization, an HTLV-I infected individual must have an antibody response to either gp46 or gp62/68 (both Env protein bands) and either p19, p24, or p53 (one of the Gag bands). Even more rigid criteria has been recommended by the HTLV European Research Network, suggesting bands for both Gag proteins p19 and p24 as well as the Env proteins rgp21 and rgp46-I must be present to ensure seropositivity [15].


Figure 1.
The process of identifying and classifying seroindeterminate, negative, and positive Human T-lymphotropic virus type I (HTLV-I) samples by enzyme immunoassay (EIA) testing and confirmatory Western blot (WB).
Generally, HTLV-I infected individuals such as asymptomatic carriers, HAM/TSP patients, and ATL patients exhibit the clear seropositivity illustrated in Figure 2 on the HTLV blot 2.4 (MP Diagnostics, Solon, OH, USA). However, there have been reports around the world of HTLV-I/II seroindeterminate WB banding patterns that do not meet the strict criteria of HTLV-I seropositivity [1521]. These are referred to as HTLV-I/II seroindeterminates (see Figure 2, blots IND-1, IND-2, and IND-3). HTLV-I/II seroindeterminate samples have been identified in otherwise healthy and normal individuals from Jamaica, Japan, Brazil, and other HTLV-I endemic areas and showed no other sign of disease or viral infection. However, of particular interest are the reports of HTLV-I/II seroindeterminate blotting patterns in patients with various neurological diseases including Multiple Sclerosis [12,22]. While the significance of HTLV-I/II seroindeterminates is still under investigation, it is clear from this finding that these samples may relate to disease (specifically neurodegenerative diseases) in at least a portion of cases. The reported prevalence of seroindeterminates has varied greatly from .023% in Taiwan, to 0.1% in Argentina, to as high as 20% of a high-risk population in Brazil [18,23,24]. Interestingly, when DNA isolated from peripheral blood mononuclear cells (PBMC) of these HTLV-I/II seroindeterminate individuals is amplified using polymerase chain reaction (PCR) assays, typically no HTLV-I or HTLV-II virus is detected (however, recent reports from Iran, Argentina, and Brazil have challenged this finding) [17]. A seroindeterminate sample is defined by a positive EIA result but an incomplete banding pattern by WB, as depicted in Figure 1 [16]. Incomplete banding patterns have manifested in various ways. A number of samples yielded all core protein bands, but lacked one or both of the recombinant bands (GD21 and rgp46-I) necessary for seropositivity (for example, IND-2 in Figure 2). Other samples seemed to exhibit a negative blotting strip except for a single protein band (as shown on IND-3 in Figure 2). Classifying a blot as seroindeterminate can vary greatly between studies and cohorts.


Figure 2.
The first two Western Blot strips depict positive control HTLV-I and HTLV-II Western Blots exhibiting all core bands necessary for seropositivity inclusion, and other bands exhibited in a typical infected sample. IND-1 IND-2 and IND-3 illustrate three ...
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3.Possible Explanations for HTLV-I/II Seroindeterminates
As research progresses, reports of large numbers of HTLV-I/II seroindeterminates have come from Iran, Brazil, Japan, Argentina, Taiwan, the Caribbean, Central Africa, and the United States, indicating a higher prevalence than originally thought [1521]. Importantly, the reasons for these blotting patterns remain unclear. Several possible explanations have been proposed for the occurrence of HTLV-I/II seroindeterminates that include cross-reactivities to other known retroviruses or a novel virus, antibody responses to a malaria parasite with epitope homology to HTLV-I, a defective HTLV-I or HTLV-II, and low copy numbers of prototypic HTLV-I in the affected patient yielding the indeterminate antibody response.

3.1. Malaria and Severe Acute Respiratory Syndrome (SARS)

It has been reported that anti-HTLV-I reactivity of sera can result from antibodies produced in response to Plasmodium falciparum (P. falciparum). P. falciparum is a lethal malaria parasite and is the most prevalent of malaria parasites infecting humans [14]. Studies regarding this cross-reactivity concluded that P. falciparum and HTLV-I must contain regions of immunogenic epitope homology. It was hypothesized that this homology may be a result of mimicry of host tissue by the two organisms [25]. This suggests that in geographic regions known to be endemic for malaria, such as the Philippines, and in which high levels of HTLV-I antibody reactivity were reported, HTLV-I/II seroindeterminates are difficult to interpret, as it is difficult to rule out the possibility of cross-reactivity between HTLV-I/II and P. falciparum [25]. It was later reported that this cross-reactivity might not be limited to P. falciparum. Examination of data from a separate cohort showed cross-reactivity with HTLV-I could be blocked by erythrocyte lysates (from areas where malaria is endemic) infected with P. falciparum, however HTLV-I seroindeterminate samples from the United States failed to block antibody reactivities. This led to the assertion that the molecular mimicry may extend to epitopes other than the P. falciparum and plasmodial antigens [26]. However, HTLV-I/II seroindeterminate banding patterns are being reported in areas where exposure to P. falciparum is extremely unlikely, such as the United States. Furthermore, HTLV-I/II seroindeterminate patterns are observed in normal, healthy blood donors, showing no sign of malaria or similar parasite infection [20]. While a subset of HTLV-I/II seroindeterminate samples may exhibit an antibody cross-reaction between HTLV-I and P. falciparum, this cannot explain the WB banding patterns found in areas where these organisms are extremely rare or nonexistent.

It should also be noted that cross-reactivity between antibodies against severe acute respiratory syndrome (SARS) coronavirus and HTLV has been observed on occasion [27]. The HTLV antibody response was exhibited when numerous serology tests were performed on samples from patients admitted to a hospital in Taiwan for SARS. However, this was immediately following the SARS outbreak in 2003, and has not been observed in other regions.

3.2. Cross-Reactivities to Other Retrovirus

Various other hypotheses have been presented regarding the cross-reactivity of anti-HTLV-I antibodies. In 1997, it was reported that HTLV I/II indeterminate serologies were observed as responses to two types of simian T-lymphotropic virus (STLV) [28]. An HTLV-I/II seroindeterminate sample that was also HTLV-I PCR positive was sequenced and showed a divergent form of HTLV-I [28]. Further investigations led to the suggestion that indeterminate blots may denote an antibody response to immunogenic regions of an endogenous retrovirus [29]. Also reported was the demonstration of elevated antibody levels to synthetic HTLV-I epitopes in samples previously testing negative for HTLV-I antibodies. The reactivity in this study reinforced a previous suggestion that increased levels of HTLV-I antibodies may be an autoantibody response to the endogenous retrovirus HRES-I [30,31]. The specific homologous regions between HTLV-I and these endogenous retroviruses occur in the LTR, gag, and pol regions of the virus. However, a later report demonstrated the tax region of prototype HTLV-I virus was amplified by nested PCR from one patient with an HTLV-I/II seroindeterminate WB that could not have been derived from the DNA sequence of an endogenous virus [12]. HTLV-I/II seroindeterminate banding patterns have also been reported in samples which were PCR positive for HTLV-I, supporting the exciting possibility that an HTLV-I/II seroindeterminate pattern may result from cross-reactivity with a novel virus such as HTLV-III or HTLV-IV [32]. These newly discovered human retroviruses were found in Cameroonese hunters showing no signs of HTLV-related diseases, and all four HTLV types show 6070% sequence homology with each other [31].

3.3. Low Copy Number of Prototype HTLV-I

Due to the typically negative PCR results and lack of antibody response to some of the HTLV-I antigens but reactivity to others, the most plausible suggestions seems to be that HTLV-I/II seroindeterminate blots may result from a low copy number of prototypic HTLV-I [12]. This explanation is supported by studies showing the ability to amplify the HTLV-I tax region from PBMCs of some HTLV-I/II seroindeterminates, but not other regions of the virus [12,28,29]. The same study reported the successful generation of an Epstein-Barr virus transformed B-cell line from a relapsing remitting multiple sclerosis patient with a seroindeterminate WB. The PBMCs from this patient had tested negative for regions of HTLV-I by PCR, while an in vitro long-term generated B-cell line tested positive by primary PCR. The virus infecting the seroindeterminate B cell line was then sequenced in an attempt to identify any mutations or other factors that may be associated with an HTLV-I/II WB seroindeterminate status. The results indicated that the virus was globally >97% homologous to prototypic HTLV-I on the nucleotide level. Fine analysis of the 5 LTR indicated that the HTLV-I strain infecting the patient was of the Cosmopolitan subtype [22]. This was the first reported verification of a PCR negative seroindeterminate sample resulting from infection of a full length HTLV-I virus [22]. Further support for the suggestion that these seroindeterminates may be the result of a low copy number of prototype HTLV-I comes from a study of patients transfused with HTLV-1 infected blood [33]. Eight seronegative individuals developed seroindeterimnate WB patterns after receiving a blood transfusion with the infected blood, further implicating the role of exposure to HTLV-I in seroindeteriminates [33].

The hypothesis that HTLV-I/II WB seroindeterminates (or subsets of these individuals) may be related to prototype HTLV-I is further supported by the recent observations that demonstrated HTLV-I PCR positive results in 12.5% of HTLV-I/II seroindeterminates from Iran [17] (Table 1). Consistent with this observation are reports from Brazil and Argentina, which also demonstrated comparable rates of PCR reactivity in HTLV-I/II seroindeterminates (9.2% and 13.8%, respectively) [17] (Table 1). The most prevalent seroindeterminate banding pattern observed in Iran was the appearance of GD21 alone, which is similar to the patterns seen in Taiwan [27,34]. The data in Table 1 also demonstrates a relatively high frequency of HTLV-I/II WB seroindeterminates from HTLV-I EIA reactive samples that are observed in multiple cohorts throughout the world. These range from 16% in the United States to 61% in Argentina. Reports of PCR amplification of prototype HTLV-I sequences from HTLV-I/II seroindeterminate individuals, especially in endemic regions such as Iran and Brazil, suggest a potentially far greater exposure to HTLV-I at least in some sub-populations of HTLV-I/II WB seroindeterminates.


Table 1.
The table illustrates the prevalence of Human T-lymphotropic virus type I (HTLV-I) in various cohorts, as well as the percentage of HTLV-I Western blot (WB) seroindeterminates in the respective cohort and the percentage of these seroindeterimates that ...
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4.Current Research and Future Directions
While the significance of these prevalent seroindeterminate HTLV-I/II blotting patterns remains uncertain, it is clear that further investigation is necessary. The sequencing of a virus with homology to prototype HTLV-I from a seroindeterminate cell line implicates exposure to the virus in at least some HTLV-I/II seroindeterminates. This finding may suggest that exposure to HTLV-I is greater than previously estimated. Of concern are reports that have demonstrated serum samples with negative HTLV-I/II results by EIA, which may also have seroindeterminate banding patterns by WB [12]. Since these sera were initially screened as HTLV-I/II EIA negative (see Figure 1), these samples (that also have an HTLV-I/II seroindeterminate WB pattern) would enter the blood supply and be available for transfusion [37]. There have also been instances of serum samples testing EIA positive and WB indeterminate at one point in time, and later testing EIA negative, while remaining seroindeterminate by WB [33]. This implies testing at one time point by EIA cannot ensure the exclusion of seroindeterminate samples. If seroindeterminate blotting patterns are a result of infection or exposure to HTLV-I (a virus associated with neurodegenerative disease, among other illnesses), and blood samples with these patterns can be negative by EIA screening and enter the blood supply, this poses a possible risk to those transfused with HTLV-I/II seroindeterminate blood although the clinical consequence of obtaining blood from an HTLV-I/II WB seroindeterminate donor is not known. Based on these observations there is a need for more stringent screening techniques for blood banks to ensure blood from seroindeterminate donors is not transfused. Increasingly sensitive and specific assays are currently being optimized to better detect anti-HTLV-I antibody responses.

One such technique is LIPS assay. The LIPS assay yields high throughput results while also generating quantitative values of anti-HTLV-I antibody responses [9]. This creates data for quantitative cohort comparison as well as allows for differentiation between HTLV-I infected disease patients and asymptomatic HTLV-I carriers and the possible generation of a risk assessment for disease development. Using antibody responses to the immunogenic epitopes of HTLV-I, Gag, Env and Tax, LIPS produces clear quantitative values for asymptomatic carriers, HAM/TSP patients, ATL patients, and may yield similar results for seroindeterminate individuals. The plasmids used in the LIPS assay are inserted with a gene including immunogenic epitope of a virus (for example, the gag, env, or tax region of HTLV-I). Renilla luciferase-fusion proteins (Ruc-antigen) are obtained from cell lysates transfected with each plasmid. A preincubated serum/antigen mixture is added to a filter plate coated with immobilized protein A/G beads. This mixture is then washed to remove any fusion proteins that did not bind to the antibody. Luciferase activity is then measured in a microplate luminometer.

The LIPS assay provides a promising high throughput method of anti-HTLV-I antibody detection. Further examination of immunogenic epitopes, such as HTLV-I Rex and HBZ, may enhance the specificity and sensitivity of the assay. Currently, a whole Gag protein is used for detection of antibody response; however, antibody responses for p19, p24, or other portion of the Gag protein may yield even more specific results. A panel of these quantitative antibody responses may be able to detect seroindeterminate samples in a fast, inexpensive, and efficient manner. LIPS may also detect seroindeterminates that tested negative by EIA, proving to be a more sensitive assay in terms of seroindeterminate detection. While the LIPS assay is under further development, other methods to improve detection and classification of seroindeterminates have also been proposed including: real-time PCR, molecular diagnostic analysis, detection of amino acid changes in the env region of HTLV-I, inclusion of other synthetic peptides in serological assays, and optimization of an enzyme oligonucleotide assay (EOA) [19,3739].

With further in depth research involving these newer methodologies, the significance of these HTLV-I/II seroindeterminate banding patterns will become clearer. As it becomes apparent that the prevalence of seroindeterminates is more widespread than originally thought, it becomes increasingly important to clarify the meaning of these results. If HTLV-I/II seroindeterminates (or subsets of these subjects) represent exposure to prototype HTLV-I or HTLV-II, the frequency of infection in the general population will be greater than previously believed. It remains to be seen whether the incidence of exposure is also associated with any clinical disease outcome.
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replied February 3rd, 2013
Experienced User
Discussion
LIPS technology provided quantitative measures of antibody titers to most of the proteins of HTLV-I. This simple modular system, expressing HTLV-I antigens as a series of Ruc fusion proteins followed by standardized chemiluminescent detection, efficiently evaluated patient humoral response to these different HTLV-I antigens. The substantial difference in anti-Gag antibody titers between the HTLV-I-infected samples and normal HTLV-I seronegative controls as determined by LIPS allowed for a universal and rigorous statistical cut-off (the mean of the 42 controls plus 5 standard deviations). Of the 7 antigens tested for diagnosing HTLV-I infection, the anti-Gag antibody test was the most informative, achieving 100% sensitivity and 100% specificity. The LIPS assay likely detects more conformational epitopes than alternative immunoassay formats providing high sensitivity, high specificity and robust signals that provide a substantial and clear distinction between positive and negative HTLV-I sera. Additionally, the high-throughput format used here makes this approach highly feasible for screening large numbers of sera samples.
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replied February 5th, 2013
Experienced User
Hey guys, just out of curiosity what is everyone's age if you don't mind me asking? I'm 35. Also is anyone experiencing knee pain and sore legs? It's like when I kneel down I have to pull myself back up with my arms I can't push myself back up with my legs anymore like I used to because they are so sore and weak now. Also behind my knees where the legs bend is sore like a numb feeling its hard to explain.
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replied February 5th, 2013
Hi Everyone,

Firstly thank you all (especially Tony) for the work you are doing regarding this research & fight for acknowledgement & ignorance regarding HTLV.
I found this site after months of devastating stress & anxiety relating to what I thought was a HIV infection. But after numerous tests & at the 4 month window testing negative to all STD's & HIV tests I've realised that HTLV must be the answer to all of my worries & ongoing symptoms.
I had all the classic initial symptoms that led me to believe I had HIV. Now after 6 months post exposure the symptoms I have are swollen lymph nodes of the neck & groin, high WBC, lump in my throat, Candida on the tongue, a yeast infection in the groin area (which I have been treating with a cream) and pins & needles in my hands.

I have made an appointment with an infectious disease doctor two weeks from now on the 26th Feb (the soonest he could see me) where I was going to ask for a PCR & antibody test, but going from all the research I have done & seeing as Tony, you have done numerous ones, all coming back negative, this may very well just be burning money due to the high false negative reports I have seen. I will enquire about a Western Blot test if that is possible instead. Tony have you managed to do a WB test for HTLV? If not is it just not available to you?

Much like most of you, my expose also happened once, I gave oral to a girl for only 15 seconds, had protected sex but here I am, utterly a wreck of a person emotionally. And to top it all off, she was Brazillian, so you can imagine my horror when I discovered where HTLV is epidemic.

I'm going to be honest, I've been in a really dark place the past few months, with thoughts of ending my life seriously coming into play. Thankfully I'm over that now, there are days when I break down but they are becoming few & far between. I just try to enjoy the simple things in life, even though this disease is the last thing I think about at night & the first thing when I wake up. If the symptoms weren't such a reminder of this mistake it would be easier to get on with my life.

And to answer your question Scared49 I'm 29 years old, but don't have any leg/back issues as yet.
Best of luck to all of you. I will update you when I discuss with my doctor regarding testing but am prepared for a huge amount.
One other point I should make is I have also been diagnosed with Crohns Disease for 7 years & have never had any of the previous symptoms associated with this autoimmune disease so something is definitely wrong & unrelated to that.

I wish you all the best & thank you for this community you have created for people like us struggling with this disease.
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