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Number7

Herpes Testing-Are they really Accurate?

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Number7

Has anyone had the experience similar to the forum post below?

http://www.herpes-coldsores.com/messageforum/showthread.php?1433-Do-Herpes-tests-work-at-all

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Blessed despite

I know you already know my story Number7 but I thought I'd post it here to share with others. I was diagnosed in June this year with a positive culture for HSV 2. I had a blood test taken at the same time that was negative (herpesselect) for HSV 1 and 2 with HSV 2 value of .11. I thought I had a UTI after coming home from a vacation with my boyfriend. We broke up at the end of the vacation and 4 days later I was visually diagnosed pending the above testing. I had not been with anyone else in 2 years. I had all the classic symptoms of a primary outbreak including itching followed by swollen lymph nodes and painful urination. I have been tested twice since with positive HSV2 values of 2.36 then most recently 3.29. I contacted my ex who has been tested three times this year all negative. The first time he was tested was due to a different illness which the herpes virus is the most common cause. I have not seen his test yet and do not know which test he had as we are not on good terms but he is supposed to show it to me soon. Ill let you know how that turns out if/when it happens.

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Number7
I know you already know my story Number7 but I thought I'd post it here to share with others. I was diagnosed in June this year with a positive culture for HSV 2. I had a blood test taken at the same time that was negative (herpesselect) for HSV 1 and 2 with HSV 2 value of .11. I thought I had a UTI after coming home from a vacation with my boyfriend. We broke up at the end of the vacation and 4 days later I was visually diagnosed pending the above testing. I had not been with anyone else in 2 years. I had all the classic symptoms of a primary outbreak including itching followed by swollen lymph nodes and painful urination. I have been tested twice since with positive HSV2 values of 2.36 then most recently 3.29. I contacted my ex who has been tested three times this year all negative. The first time he was tested was due to a different illness which the herpes virus is the most common cause. I have not seen his test yet and do not know which test he had as we are not on good terms but he is supposed to show it to me soon. Ill let you know how that turns out if/when it happens.

Thanks for responding.

Yes - I'd like to raise the awareness to all of those who have had a similar experience that way more people than anyone can imagine really have hsv2. The guy in the thread I pasted above was proactive enough to start to question his testing probably because he had kept in touch with those two partners and actually had a conscience. Most guys like yours Blessed Despite ......run.

The guy I was with has promised to let me know if "anything comes up"----sure doesn't help me knowing I have it but the saying is true.......what you don't know doesn't hurt you.

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Number7

Feel free to read the article below ---

Sexually Transmitted Diseases

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> April 2003 - Volume 30 - Issue 4 > Time Course of Seroconversion by HerpeSelect ELISA After Acq...

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Sexually Transmitted Diseases:

April 2003 - Volume 30 - Issue 4 - pp 310-314

Articles

Time Course of Seroconversion by HerpeSelect ELISA After Acquisition of Genital Herpes Simplex Virus Type 1 (HSV-1) or HSV-2

ASHLEY-MORROW, RHODA PhD*; KRANTZ, ELIZABETH MS*; WALD, ANNA MD*†

Free Access

Article Outline

Author Information

*Department of Laboratory Medicine and †Departments of Medicine and Epidemiology, University of Washington, Seattle, Washington

Supported by a grant from the National Institutes of Health (no. AI 30731).

Reprint requests: Rhoda Ashley-Morrow, PhD, Childrens' Hospital and Regional Medical Center, Room G815, Virology Office, 4800 Sand Point Way NE, Seattle, WA 98115. E-mail: rashle@chmc.org

Received June 18, 2002,

revised August 28, 2002, and accepted September 23, 2002.

Abstract

Background: HerpeSelect HSV-1 and HSV-2 ELISAs are glycoprotein G-based, type-specific antibody detection tests that are approved by the US Food and Drug Administration for diagnosis of genital herpes.

Goal: The goal was to determine seroconversion times by means of HerpeSelect ELISAs.

Study Design: Four-hundred thirteen sera from 113 patients with recently acquired genital herpes were tested by HerpeSelect ELISAs and Western blot (WB). Thirty-one patients had primary genital HSV-1 (group 1), 56 had primary HSV-2 (group 2), and 26 had prior HSV-1 antibodies and newly acquired HSV-2 (group 3).

Results: Median interval from onset of symptoms to seroconversion was 25 days, as determined by HerpeSelect HSV-1, versus 33 days by WB for group 1; 21 days by HerpeSelect HSV-2 versus 40 days by WB (group 2;P = 0.0005); and 23 days by HerpeSelect HSV-2 ELISA versus 47 days by WB (group 3;P = 0.02). In long-term follow-up, transient reversion to HerpeSelect negativity occurred in 3 of 31 HSV-1-infected subjects (10%) and in 2 of 82 HSV-2-infected subjects (2%).

Conclusion: Seroconversion to HSV-2 was determined faster by HerpeSelect than by WB.

GENITAL HERPES INFECTIONS can be difficult to diagnose on the basis of clinical criteria alone. 1,2 Testing genital lesions by viral culture or antigen detection methods, while valuable for definitive and specific diagnosis of herpes simplex virus (HSV) infection, is insensitive. 3,4 The protean presentations of genital herpes and the insensitivity of viral detection tests can, in part, explain the underdiagnosis of genital herpes infections. 5,6 Antibody tests are especially useful to diagnose recurrent genital symptoms in patients who are not positive virologically, such as with culture or viral antigen tests. 7,8 Serology is also of value for patients without symptoms but with history of exposure to genital herpes. 9,10 Some experts believe that determining HSV serostatus is an important component of prenatal care because most neonatal herpes infections are transmitted by women who are unaware of their infections. 11

Only tests based on the HSV type-specific protein glycoprotein gG can accurately differentiate between antibodies to HSV-1 and those directed to HSV-2. 12-14 Only two commercial tests, HerpeSelect ELISA and HerpeSelect Immunoblot (Focus Technologies, Cypress, CA) are approved by the US Food and Drug Administration (FDA) for detection of HSV-1 as well as HSV-2 type-specific antibodies. While both of these tests are intended for laboratory testing of sera, the ELISA format is less expensive and more amenable to large-scale testing. A third FDA-approved gG-based test, POCkit-HSV-2 (Diagnology), is a point-of-care membrane test for antibodies to HSV-2. 15,16

Western blot (WB) has been used to describe, in detail, the kinetics of seroconversion to HSV-1 and HSV-2 and the relative times required to develop antibodies to individual HSV proteins. 13,17 In one series of patients, WB detected 92% of new HSV-1 infections and 93% of new HSV-2 infections within 6 weeks of symptom onset. 18 However, WB is expensive to perform and is not widely used. This study was designed to establish the time to seroconversion, as determined by HerpeSelect HSV-1 ELISA and HerpeSelect HSV-2 ELISA, in a characterized cohort of patients with newly acquired, culture-documented genital HSV-1 infections (n = 31) or HSV-2 infections (n = 82). The data obtained were compared with results of WB testing.

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Materials and Methods

Subjects

Patients were enrolled at the University of Washington Virology Research Clinic between May 1981 and December 2000 for studies of the natural history and epidemiology of genital herpes. Informed consent was obtained to store sera for testing for HSV antibodies. Eligibility criteria for this study were as follows: (1) at least one culture (of a specimen from an anogenital site) positive for HSV-1 or HSV-2 during the first clinical episode of genital herpes; (2) documented new acquisition of genital HSV infection at the time of presentation to the clinic, as defined by lack of a full antibody profile (per WB) to the same type as the culture obtained at the first visit; (3) a maximum of 28 days from the first symptoms to the first visit at which serum was drawn; and (4) at least one follow-up clinic visit and serum sampling within 180 days of the first visit.

At the first visit, a standardized questionnaire was administered to elicit information about the first symptoms of genital herpes. A physical examination was performed and all genital lesions were recorded. Women underwent a speculum examination of the cervix and vaginal vault. Samples for herpes culture and virus typing were obtained by swabbing lesions. Separate swabs were used to sample the cervix, vulva, perineum, and perianal areas of women or from penile skin, perineum, and perianal areas of men. 19 Blood was obtained for HSV serology. Subjects returned to the clinic for repeated testing during recurrent episodes of genital symptoms and-depending on study protocol-at additional times to provide information on the natural history of genital herpes. Subjects were not known to be HIV-seropositive during the period of time that sera were drawn.

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Serologic Testing

Enrollment sera were paired with convalescent sera obtained 6 to 8 weeks later. WB was performed to confirm patients' infection category as primary (lack of full antibody profile to either type at the first visit) or nonprimary HSV-2 (antibodies only to HSV-1) at the first visit. 18

For this study, sera from a single patient were assembled into chronologic sets from the serum bank and thawed, and two separate aliquots were prepared for WB (aliquot 1) or HerpeSelect HSV-1 and HSV-2 ELISAs (aliquot 2). WB profiles were scored as negative (no HSV-specific bands), limited (fewer than four HSV-specific bands), or positive (four or more HSV-specific bands, including gG-2 for sera positive for HSV-2). The results of HerpeSelect HSV-1 and HSV-2 ELISAs (purchased from Focus Technologies) were recorded as follows: index values <0.9, negative; >1.1, positive; and inclusive values between 0.9 and 1.1, equivocal. Seroreversion was defined by a positive ELISA result followed later by a negative result.

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Statistical Methods

Time to seroconversion was defined as the number of days from the onset of primary symptoms (day 0) to the first positive serological test result, for each respective test. Subjects for whom no test results were positive were censored at their last visit. To maintain an acceptable level of accuracy in estimating time to seroconversion, subjects who had gaps of greater than 180 days between sera collections were censored at the last visit before such a gap occurred. Twenty-five of the 113 subjects had gaps greater than 180 days between two sera collections, but most of these subjects (20 of 25) had seroconverted before the gap in sampling occurred. Kaplan-Meier curves were constructed to estimate median times to seroconversion.

Log rank tests were used to test for differences in time to seroconversion between independent groups defined by clinical category, whereas paired Prentice-Wilcoxon tests were used to assess the differences between time to seroconversion for different serologic tests on the same subjects within a clinical category. 20

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Results

Description of Data

The length of time between a subject's first symptoms and the first visit to the clinic ranged from 0 days (the subject reported to the clinic the first day symptoms were noted) to 28 days (median, 4 days). Follow-up time for subjects in this study was intended to simulate the clinical scenario of a patient having blood drawn at the acute phase of illness and then during the convalescent phase, several weeks to several months later. Patients with primary HSV-1 infection (group 1) were followed a median of 89 days, those with primary HSV-2 infection (group 2) for a median of 93 days, and those with nonprimary first episodes of HSV-2 infection (group 3) for a median of 109 days. Follow-up occurred within the first year of infection. Patients in group 1 had a median of 3 visits per person (range, 2-10); those in group 2 had a median of 3 visits per person (range, 2-7); and those in group 3 had a median of 4 visits per person (range, 2-9). No subject was tested beyond a year after onset of the primary or nonprimary first episode.

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Time to Seroconversion by HerpeSelect

Figure 1 shows survival curves indicating the time to seroconversion for each of the three study groups. Median time to seroconversion by HerpeSelect was 25 days for group 1 (primary HSV-1;Figure 1A), 21 days for group 2 (primary HSV-2;Figure 1B), and 23 days for group 3 (nonprimary HSV-2;Figure 1C). Sixty-two percent of subjects infected with HSV-1 (group 1), compared with 59% of subjects infected with HSV-2 (group 2), seroconverted by 6 weeks. Seventy-seven percent of patients with nonprimary HSV-2 infection (group 3) seroconverted by 6 weeks (Table 1). Time to seroconversion, as determined by HerpeSelect, did not differ significantly among the three groups (P = 0.16; log rank test).

Fig. 1

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Table 1

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Time to Seroconversion by WB

Median time to development of a full WB profile was 33 days for group 1 (Figure 1A), 40 days for group 2 (Figure 1B), and 47 days for group 3 (Figure 1C). Sixty percent of subjects with primary HSV-1 infection had full WB profiles to HSV-1 by 6 weeks. Full HSV-2 profiles developed by 6 weeks for 36% to 53% in groups 3 and 2, respectively (Table 1). Time to seroconversion, as determined by WB, did not differ among the three groups (P = 0.22; log rank test).

Positivity of a WB test is based on the development of antibody responses sufficient to show at least four bands. 18 However, seroconversion is a dynamic process, with antibodies to different HSV proteins developing at different rates. 17,21 Thus, when a progressive change from a seronegative blot to one with fewer than four specific bands is noted in paired testing of at least two sera, this limited profile can be highly predictive of recent HSV infection. 22 Therefore, we examined time to seroconversion on the basis of the first indication of a limited profile to the appropriate HSV type. Not surprisingly, median times were less than those required for a full WB profile to develop: 29 days for group 1 (vs. 33 days for a full HSV-1 profile), 19 days for group 2 (vs. 40 days for a full HSV-2 profile), and 34 days for group 3 (vs. 47 days for a full HSV-2 profile) (Figure 1A-C). A higher proportion showed limited WB profiles than full WB profiles at 6 weeks and 3 months (Table 1).

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Differences Between Tests of Time to Seroconversion

Time to seroconversion by HerpeSelect versus time to development of a full WB profile was compared by the paired Prentice-Wilcoxon test. 20 No statistically significant difference in time to HSV-1 seroconversion was seen (group 1;P = 0.12). However, HSV-2 seroconversion was determined significantly faster by the HerpeSelect test than by WB (P < 0.001 for group 2 and P = 0.02 for group 3).

When HerpeSelect results were compared with results based on observing a limited WB profile, the WB was faster in demonstrating primary HSV-2 seroconversion (group 2;P = 0.01). However, no significant differences between the two tests were seen for subjects with primary HSV-1 infection (P = 0.45) or subjects with nonprimary first episodes of HSV-2 infection (P = 0.23).

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Subjects Without Documented Seroconversion

Overall, 15% of subjects (17 of 113) did not show seroconversion by HerpeSelect during the study period: 7 of 31 (23%) in the HSV-1 primary group, 7 of 56 (13%) in the HSV-2 primary group, and 3 of 26 (12%) in the nonprimary HSV-2 group. The 17 HerpeSelect nonseroconverters were followed a median of 54 days (range, 15-208) from the date of primary onset, compared with a median of 97 days (range, 14-361) for subjects who did seroconvert.

Sixteen of 113 (14.2%) did not seroconvert by WB. The median time of follow-up for the 16 WB nonseroconverters was 37 days (range, 15-167), compared with 98 days (range, 14-361) for subjects who seroconverted. Among 89 subjects who seroconverted by both tests, the median time for follow-up was 99 days (range, 14-361).

Nine of 113 subjects-3 infected with HSV-1, 3 with HSV-2, and 3 with nonprimary HSV-2-did not seroconvert by either test (median follow-up time, 38 days; range, 15-167). Thus, for both HerpeSelect and WB tests, apparent failure to seroconvert may have been a function of short follow-up time.

Some patients failed to seroconvert by HerpeSelect but became WB-positive, whereas others seroconverted only by HerpeSelect. Of those with primary HSV-1 infection (group 1), 4 seroconverted by WB but not by HerpeSelect, whereas 2 seroconverted by HerpeSelect but not by WB; for those with primary HSV-2 infection (group 2), 4 showed seroconversion by WB but not by HerpeSelect, and another 4 seroconverted by HerpeSelect but not by WB. One subject with nonprimary HSV-2 infection seroconverted by HerpeSelect but not by WB.

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Increasing Sensitivity by Combining Tests

Because of the differences in seroconversion times and prevalence between the two tests, we explored the use of combined HerpeSelect and WB results for the most effective identification of new infections. Using a definition of seroconversion time as the minimum observed time to a positive HerpeSelect test or an indication of a change in the WB profile or to a full WB profile, we found that only 1 (3%) of 31 subjects with primary HSV-1 infection, 3 (12%) of 26 subjects with nonprimary HSV-2 infection, and no patients with primary HSV-2 infection failed to seroconvert by any measure. Median time to seroconversion was reduced by about a week in each group when earliest time to positivity by either WB or HerpeSelect was compared with time to seroconversion by only HerpeSelect: 17 versus 25 days for primary HSV-1 infection, 17 versus 21 days for primary HSV-2 infection, and 13 versus 23 days for nonprimary first episodes of HSV-2 infection. The proportion of patients seroconverting by any test by 6 weeks was 88% for group 1, 93% for group 2, and 71% for group 3 (Table 1).

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Seroreversion

gG-based Tests -Some studies have indicted that tests could become falsely negative over time for men 23 and over the course of pregnancy for women. 24 While our study was designed to determine time to seroconversion in sera drawn early in the course of infection, we tested sera for as long as a year after onset of genital herpes to demonstrate seroconversion in patients who might require longer to become positive by either HerpeSelect or WB tests. We examined all follow-up sera for this analysis, including sera drawn more than 180 days apart and, for subjects appearing to serorevert, sera drawn more than 1 year later if available.

Of 31 primary HSV-1 patients, 3 (10%) became positive by HSV-1 ELISA and then had at least one HerpeSelect-negative result. One subject became negative on day 22 and remained negative in tests of eight samples drawn between days 22 and 376. However, WB became positive on day 8 and remained positive throughout follow-up for this subject. Another subject became positive by HSV-1 ELISA at 17 days but was negative in testing of a single serum drawn on day 158. WB was negative on days 17 and 158 for this subject. The third subject became positive on day 70 by HSV-1 ELISA and then was transiently negative in testing of one serum on day 372, before returning to positive on day 387. WB for this subject became positive on day 29 and remained positive.

Of 82 patients with genital HSV-2, two (2%) had negative test results following at least one positive HSV-2 result by HerpeSelect. One subject became positive on day 9, was negative on day 81, and was subsequently seropositive on days 91, 101, 168, 191, and 361. WB became positive for HSV-2 on day 91 for this patient and did not change thereafter. The second patient became positive on day 14 and then had an equivocal result on day 14 and a negative result on day 108. WB became positive for HSV-2 on day 14 and remained positive.

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Discussion

The FDA has approved three gG-based tests for diagnosing HSV infections: Focus Technologies' HerpeSelect ELISA for HSV-1 and HSV-2 antibodies, HerpeSelect immunoblot for HSV-1 and HSV-2 antibodies, and Diagnology's POCkit-HSV-2 for HSV-2 antibodies. In addition, gG-based ELISAs from Gull Laboratories were FDA-approved for diagnosis of HSV-1 and HSV-2 infection and marketed briefly by Meridian Biosciences under the Premier brand name. 25 The Gull/Meridian tests are no longer available. WB testing is considered the gold standard for detecting HSV antibodies, but it is not FDA-approved and is not widely available.

Of these tests, HerpeSelect ELISA is the most cost-effective test for commercial laboratory testing. Published data on HerpeSelect ELISA indicate a sensitivity of 98% and specificity of 94% for HSV-1 antibodies 26 and sensitivity of 98% and specificity of 95% for HSV-2 antibodies. 27 However, sensitivity may be lower in newly infected persons who have not developed high antibody titers. Therefore, we selected 113 patients with culture-documented first episodes of genital herpes to test in parallel by HerpeSelect ELISA and WB to determine time to seroconversion. Median time from onset of symptoms to seroconversion by HerpeSelect ELISA was 25 days for patients with primary genital HSV-1 infection, 21 days for those with primary HSV-2 infection, and 23 days for those with prior HSV-1 antibodies and new (nonprimary) genital HSV-2 infections.

HerpeSelect was significantly faster for HSV-2 seroconversion than was development of full antibody profiles by WB. However, the time required for development of the first evidence of de novo, HSV-specific bands (a less stringent criterion than a full antibody profile) was significantly less than with HerpeSelect for patients with HSV-2 primary infection.

The practical applications of our study relate to diagnosis of suspected new genital herpes infections. A positive HerpeSelect test within 6 weeks of lesion onset is probably an accurate reflection of genital herpes. However, a negative test result occurred in our study for 38% of HSV-1-infected patients in the first 6 weeks and in 23% to 41% of HSV-2-infected patients. By 3 months, between 73% and 93% of patients had seroconverted to either HSV-1 or HSV-2 by HerpeSelect. Thus, retesting at a later date appears to help detect seroconversion. Alternatively, our data show that performing WB in addition to HerpeSelect can decrease the time to detection of seroconversion for some patients. A serum found to be negative or equivocal by HerpeSelect had a full WB profile in 37% of samples from HSV-1-infected patients and in 17% of samples from HSV-2-infected patients.

Our study did not test the sensitivity of HerpeSelect for patients with subtle or subclinical infections, and it is unclear whether such patients have the same kinetics of seroconversion to gG that we recorded for symptomatic patients. Up to 90% of patients with HSV-2 antibodies do not have a history of genital herpes 5; thus, type-specific serology is an important option to diagnose unrecognized recurrent herpes. 28,29 Studies are in progress to test the sensitivity and specificity of HerpeSelect for HSV-2 infections identified by WB and subsequently confirmed when shedding of virus from the genital tract is determined by culture or PCR. 30

It should be emphasized that the current data reflect only one aspect of test performance: time to seroconversion in the patient with symptoms severe enough to result in a clinic visit. Furthermore, because our study was based on infected patients (and thus lacks positive test findings for uninfected patients), we do not have data to identify the specificity of the HerpeSelect tests.

The occurrence of seroreversion to gG has been noted for other gG-based tests. 23,24 Our study was not specifically designed to determine the frequency or clinical correlates for HerpeSelect seroreversion. However, we identified transient reversion to negative test outcomes in 3 of 31 patients with primary HSV-1 infection and in 2 of 82 patients with nonprimary HSV-2 infection. WB profiles from seroreverting patients did not change over time. Although this observation suggests that decreases in gG antibody titer were not the major factor in seroreversion, WB is not a quantitative test, and the possibility of antibody titer changes cannot be definitely dismissed. The issue for most clinicians is not the possibility of seroreversion in a patient with diagnosed infection but what the observation of seroreversion in longitudinal studies means for the detection of antibodies in patients with no such diagnosis. Larger studies to test the sensitivity of HerpeSelect over longer periods of time would be helpful.

In summary, HerpeSelect ELISAs can detect seroconversion to HSV-1 and HSV-2 within a month of symptom onset in the majority of patients. Although careful correlation of laboratory results with history and clinical observation is always important, the availability of these tests should provide clinicians with an important diagnostic tool.

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References

epidemiology of genital infection with herpes simplex virus types 1 and 2

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      @WilsoInAus 

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      THIS IS A SUPPORT COMMUNITY NOT A BASH COMMUNITY. YOU THINK IT'S OKAY TO BASH PEOPLE FOR LORD KNOW WHAT REASON.  You attempt to manipulate my character with your slander of my character.  The only person proving them self of poor character here is yourself.  

      I have reported you. You have no reason to be here, you are the classical keyboard warrior-troll. I have every right to be pissed off and trust me if I could. I would beat the shit out of you, bc you need lessons learned.  You're alien trying to tell another human being his/her symptoms and feelings.  FUCK OFF. I will never reply to you again. I don't care how much you try to stir the pot. You have lost all credibility and you have nothing better to do than try to demoralize other people. You're a shitty human being. 
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      You can have HIV for many years without developing AIDs, if that is what you're asking.
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