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Biomedical subjects

R L Ashley

Publications and source records attributed to R L Ashley.

At least 37 records · Page 2Linked to original sources
PubMed1998

Premarket evaluation of a commercial glycoprotein G-based enzyme immunoassay for herpes simplex virus type-specific antibodies.

A new commercial glycoprotein G-based enzyme immunoassay (gG-EIA) was compared with Western blotting (WB) for detection of herpes simplex virus type 1 (HSV-1) or HSV-2 type-specific antibodies in 193 serum samples. Sensitivity for HSV-1 was 95%; specificity was 96%. Sensitivity for HSV-2 was 98%; specificity was 97%. Twelve of 13 serum samples with equivocal gG-EIA results were serotyped by WB.

Antibodies, Viral↗
PubMed1998

Occult systemic infection and persistent simian immunodeficiency virus (SIV)-specific CD4(+)-T-cell proliferative responses in rhesus macaques that were transiently viremic after intravaginal inoculation of SIV.

The intact cervicovaginal mucosa is a relative barrier to the sexual transmission of human immunodeficiency virus type 1 (HIV-1). In the simian immunodeficiency virus (SIV) macaque model of HIV infection, seronegative transient viremia (STV; virus isolation positive followed by repeated negative cultures) occurs after intravaginal inoculation of a low dose of pathogenic SIVmac251 (C. J. Miller, M. Marthas, J. Torten, N. Alexander, J. Moore, G. Doncel, and A. Hendrickx, J. Virol. 68:6391-6400, 1994). Thirty-one adult female macaques that had been inoculated intravaginally with pathogenic SIVmac251 became transiently viremic. One monkey that had been culture negative for a year after SIV inoculation became persistently viremic and developed simian AIDS. No other STV monkey developed persistent viremia or disease. Results of very sensitive assays showed that 6 of 31 monkeys had weak SIV-specific antibody responses. SIV-specific antibodies were not detected in the cervicovaginal secretions of 10 STV monkeys examined. Twenty of 26 monkeys had lymphocyte proliferative responses to p55(gag) and/or gp130(env) antigens; 3 of 6 animals, including the monkey that became persistently viremic, had detectable cytotoxic T-lymphocyte (CTL) responses to SIV. At necropsy, lymphoid tissues and vaginal mucosa were virus culture negative, but in 10 of 10 animals, SIV provirus was detected by PCR using gag-specific primer pairs. Fifty percent of the PCR-positive tissue samples were also positive for SIV gag RNA by reverse transcriptase PCR. Thus, transient viremia following intravaginal inoculation of pathogenic SIV is associated with persistent, systemic infection, either latent or very low level productive. Atypical immune responses, characterized by lymphocyte proliferation and some CTL responses in the absence of conventionally detectable antibodies, develop in transiently viremic monkeys.

Animals↗
PubMed1997

The acquisition of herpes simplex virus during pregnancy.

BACKGROUND: The acquisition of genital herpes during pregnancy has been associated with spontaneous abortion, prematurity, and congenital and neonatal herpes. The frequency of seroconversion, maternal symptoms of the disease, and the timing of its greatest effect on the outcome of pregnancy have not been systematically studied. METHODS: We studied 7046 pregnant women whom serologic tests showed to be at risk for herpes simplex virus (HSV) infection. Serum samples obtained at the first prenatal visit, at approximately 16 and 24 weeks, and during labor were tested for antibodies to HSV types 1 and 2 (HSV-1 and HSV-2) by the Western blot assay, and the results were correlated with the occurrence of antenatal genital infections. RESULTS: Ninety-four of the women became seropositive for HSV; 34 of the 94 women (36 percent) had symptoms consistent with herpes infection. Women who were initially seronegative for both HSV-1 and HSV-2 had an estimated chance of seroconversion for either virus of 3.7 percent; those who were initially seropositive only for HSV-1 had an estimated chance of HSV-2 seroconversion of 1.7 percent; and those who were initially HSV-2-seropositive had an estimated chance of zero for acquiring HSV-1 infection. Among the 60 of the 94 pregnancies for which the time of acquisition of HSV infection was known, 30 percent of the infections occurred in the first trimester, 30 percent in the second, and 40 percent in the third. HSV seroconversion completed by the time of labor was not associated with an increase in neonatal morbidity or with any cases of congenital herpes infection. However, among the infants born to nine women who acquired genital HSV infection shortly before labor, neonatal HSV infection occurred in four infants, of whom one died. CONCLUSIONS: Two percent or more of susceptible women acquire HSV infection during pregnancy. Acquisition of infection with seroconversion completed before labor does not appear to affect the outcome of pregnancy, but infection acquired near the time of labor is associated with neonatal herpes and perinatal morbidity.

Adult↗
PubMed1997

Comparative analysis of methods for collection and measurement of immunoglobulins in cervical and vaginal secretions of women.

The purpose of this study was to systematically compare 3 collection methods, Sno-strips, wicks and cervical-vaginal lavage, for analysis of immunoglobulin concentrations in female genital secretions. In each of 8 women, absorbent wicks and Sno-strips were applied at 4 locations: the lateral wall of the vagina; the posterior vaginal fornix; the surface of the exocervix; and the endocervical canal. Cervical-vaginal lavage was then performed in 4 women with 5 ml PBS. Immunoglobulin and protein concentrations in lavage samples were generally over 100 times lower than in the secretions captured directly from mucosal surfaces with either Sno-strips or wicks. Capture of undiluted secretions with either wicks or Sno-strips allowed calculation of actual immunoglobulin concentrations at specific mucosal sites: for example, median IgA levels were consistently highest in the endocervix and lowest in the vagina. Such information may be crucial in evaluating the correlates of protective immunity against micro-organisms that infect or invade discrete regions of the genital mucosa.

Adolescent↗
PubMed1997

Herpes simplex virus type 2 detection by culture and polymerase chain reaction and relationship to genital symptoms and cervical antibody status during the third trimester of pregnancy.

OBJECTIVES: Our goal was to define the frequency of asymptomatic herpes simplex virus type 2 shedding by culture and polymerase chain reaction and to correlate our findings with cervical anti-herpes simplex virus type 2 immunoglobulin A production. STUDY DESIGN: Women who were seropositive for herpes simplex virus type 2 collected daily genital tract samples during the third trimester for culture and deoxyribonucleic acid quantitation by polymerase chain reaction. Cervical secretions were collected weekly for anti-herpes simplex virus type 2 immunoglobulin A. Asymptomatic shedding by culture versus polymerase chain reaction and anti-herpes simplex virus type 2 immunoglobulin A detection with and without genital shedding were compared by means of McNemar's chi 2 test. RESULTS: Asymptomatic shedding was more frequent by polymerase chain reaction than by culture (13.8% vs 2.3%, p < 0.0001). When cervical anti-herpes simplex virus type 2 immunoglobulin A was present, patients were more likely to have negative results by polymerase chain reaction than positive results (66.7% vs 26.7%, p = 0.001). Anti-herpes simplex virus type 2 immunoglobulin A was detected beyond 37 weeks in only one subject. CONCLUSIONS: Polymerase chain reaction was more sensitive than culture for detecting asymptomatic genital herpes simplex virus. The role of immunoglobulin A in clearing genital herpes simplex virus remains to be determined.

Adult↗
PubMed1997

A novel method to assay herpes simplex virus neutralizing antibodies using BHKICP6LacZ-5 (ELVIS) cells.

A novel method for determining neutralizing serum antibody titers to herpes simplex virus type 2 (HSV-2) was developed based on reduction of infectivity in BHKICP6LacZ-5 (ELVIS) cells; baby hamster kidney (BHK) cells that have been genetically engineered to contain the Escherichia coli LacZ gene under the control of an inducible herpes simplex virus type 1 (HSV-1) promoter. The test has a semiautomated, colorimetric readout resulting in rapid, objective readings of infectivity reduction. Extent of neutralization is calculated against a calibration curve of virus infectivity generated in each run. HSV-2 neutralizing activity can be detected with serum dilutions in excess of 1:5120.

Animals↗
PubMed1997

Genital herpes in a primary care clinic. Demographic and sexual correlates of herpes simplex type 2 infections.

BACKGROUND AND OBJECTIVES: Genital herpes remains one of the most prevalent sexually transmitted diseases (STDs). The sexual behavioral correlates of herpes simplex virus type 2 (HSV-2) infection in the general population have not been well characterized. GOALS: To assess demographic and sexual behavioral correlates of symptomatic and subclinical HSV-2 infection. STUDY DESIGN: Cross-sectional survey of 922 randomly chosen patients and 78 of their partners (1,000 total) in a family practice. Sexual behavior information was collected in 492 people. RESULTS: Two hundred twenty-five (23%) heterosexual people had HSV-2 infection, but only 59 (26%) reported a history of genital herpes. HSV-2 seroprevalence was 63% in African-American women, 27% in white women, 40% in African-American men, and 12% in white men. In multivariate analyses of risk factors for HSV-2 infection among men, 10 or more sexual partners and a prior STD were statistically significantly associated with HSV-2 infection. Among white women, number of sexual partners, a prior STD, marriage or cohabitation, and less than a college education were predictive of HSV-2 infection. A history of oral sex was the only statistically significant predictor of HSV-2 infection in African-American women. CONCLUSIONS: Risk factors for HSV-2 infection differ by gender and ethnic group. Traditionally recognized behavioral correlates of STD acquisition may not identify people in communities with high prevalence of HSV-2 infection.

Adolescent↗
PubMed1996

Viral serologies in patients with chronic fatigue and chronic fatigue syndrome.

Chronic fatigue syndrome (CFS) is an illness characterized by disabling fatigue associated with complaints of fevers, sore throat, myalgia, lymphadenopathy, sleep disturbances, neurocognitive difficulties, and depression. A striking feature of CFS is its sudden onset following an acute, presumably viral, illness and the subsequent recurrent "flu-like" symptoms. It has been speculated that both CFS and debilitating chronic fatigue (CF) that does not meet strict criteria for CFS may be the direct or indirect result of viral infections. We therefore tested 548 chronically fatigued patients who underwent a comprehensive medical and psychiatric evaluation for antibodies to 13 viruses. Our objectives were to compare the seroprevalence and/or geometric mean titer (GMT) of antibodies to herpes simplex virus 1 and 2, rubella, adenovirus, human herpesvirus 6, Epstein-Barr virus, cytomegalovirus, and Cox-sackie B virus, types 1-6 in patients with CF to healthy control subjects. Other goals were to determine if greater rates of seropositivity or higher GMTs occurred among subsets of patients with CFS, fibromyalgia, psychiatric disorders, a self-reported illness onset with a viral syndrome, and a documented temperature > 37 degrees C on physical examination. Differences in the seroprevalence or GMTs of antibodies to 13 viruses were not consistently found in those with CF compared with control subjects, or in any subsets of patients including those with CFS, an acute onset of illness, or a documented fever. These particular viral serologies were not useful in evaluating patients presenting with CF.

Adenoviruses, Human↗
PubMed1996

Gender differences in sexual practices and sexually transmitted infections among adults in Lima, Peru.

OBJECTIVES: This study examined the prevalences of antibodies to Treponema pallidum, Chlamydia trachomatis, and herpes simplex virus type 2 in a sample of Peruvian adults. METHODS: Among adults seeking health certification in Lima, Peru, 600 were randomly selected to undergo interviews and serologic testing. RESULTS: Men's reported mean lifetime number of partners (10.6) far exceeded women's (1.1), yet antibody to sexually transmitted infection pathogens among sexually experienced participants was 2.8 times more prevalent among women than among men. Among men, female sex workers accounted for 37% of recent partners, and only sex with female sex workers while using condoms less than half of the time was independently associated with antibody (odds ratio = 3.6, 95% confidence interval = 1.5, 8.8). among women, number of partners was associated with any sexually transmitted infection antibody, while intercourse before 18 years of age was associated with C trachomatis antibody. At every level of perceived risk, sexually transmitted infection antibody was more frequent among women. CONCLUSIONS: Men having unprotected sex with female sex workers had the greatest risk of acquiring infections and (by inference) of transmitting them to women.

Adult↗
PubMed1995

Virologic characteristics of subclinical and symptomatic genital herpes infections.

BACKGROUND: The frequency, pattern, and anatomical sites of subclinical shedding of herpes simplex virus (HSV) in the genital tract, along with factors that predict such shedding, have not been well characterized. METHODS: We studied prospectively the clinical and virologic course of genital herpes in 110 women. The women kept symptom diaries and provided daily samples from the vulva, cervix, and rectum for viral culture. RESULTS: During a median follow-up of 105 days, subclinical shedding of virus was identified in 36 of 65 women (55 percent) with HSV type 2 (HSV-2), in 16 of 31 women (52 percent) with HSV type 1 (HSV-1) and HSV-2, and in 4 of 14 women (29 percent) with only HSV-1. Among women with genital HSV-2 infection, subclinical shedding occurred on a mean of 2 percent of the days. The mean duration of viral shedding during subclinical episodes was 1.5 days, as compared with 1.8 days during symptomatic episodes. HSV was isolated from several sites in the genital tract and rectum in 17 percent of subclinical episodes and 22 percent of symptomatic episodes. Half the episodes of subclinical shedding of HSV occurred within seven days of a symptomatic recurrence. The risk of subclinical shedding increased with the frequency of symptomatic recurrences. Subclinical shedding was more frequent among women with more than 12 recurrences per year than among those with no symptomatic recurrences (odds ratio, 3.3; 95 percent confidence interval, 1.4 to 7.9); it was also more frequent among women who had recently acquired genital herpes (odds ratio for women with HSV acquired in the past year as compared with those who had had the infection for a year or more, 1.85; 95 percent confidence interval, 1.1 to 3.1). CONCLUSIONS: Among women with a history of genital herpes infection, subclinical shedding of HSV is common and accounts for nearly one third of the total days of reactivation of HSV infection in the genital tract. Women with frequent symptomatic recurrences also have frequent subclinical shedding and may be at high risk for transmitting HSV.

Adolescent↗
PubMed1995

A comparison between detailed and simple histories in the diagnosis of genital herpes complicating pregnancy.

OBJECTIVE: All women seropositive for herpes simplex virus-2 are at risk for asymptomatic viral shedding at the onset of labor and neonatal transmission of the virus. Unfortunately, only 20% to 35% of seropositive adults give a history consistent with genital herpes. We evaluated whether more detailed questioning during pregnancy might elucidate symptoms predictive of seropositivity and therefore better identify women at risk for herpes simplex virus shedding at delivery. STUDY DESIGN: During pregnancy 201 randomly selected women were asked in-depth questions about recurrent genital symptoms and whether they "currently have or have ever had genital herpes." An assessment was made whether the patient had a history compatible with genital herpes. This assessment and the "yes/no" history of genital herpes obtained by physicians at the initial prenatal visit were both compared with herpes simplex virus serologic studies by Western blot. RESULTS: Of 201 patients interviewed, 177 gave no history of genital herpes. Of these, 30.4% were seropositive for herpes simplex virus-2. Detailed histories on these 177 patients indicated that among the 159 subjects without suggestive symptoms or with somewhat suggestive symptoms the corresponding rates of seropositivity were 28% and 30%. Among the 18 (10.2%) subjects with highly suggestive symptoms, only 50% were seropositive. The positive predictive values for recurrent genital symptoms to predict herpes simplex virus-2 seropositivity ranged from 30% to 57%. CONCLUSION: A detailed history of genital symptoms is no better at identifying an herpes simplex virus-2 seropositive patient than is simply asking if she has ever had genital herpes. Serologic screening is a more accurate method of identifying women with past genital herpes or those who are at risk for acquiring genital herpes during pregnancy.

Adolescent↗
PubMed1995

Monoclonal antibody blocking tests for the detection of HSV-1- and HSV-2-specific humoral responses: comparison with western blot assay.

Monoclonal antibody blocking radioimmunoassays (MAb block RIAs) which detect specific humoral responses to each of the two human herpes simplex virus (HSV) types are described. RIAs were compared with Western blot assay (WBA) in a blind study of 64 sera obtained from clinically well-documented cases of genital herpes. WBA and MAb block RIA each detected HSV-1 antibodies in 16/17 (94%) sera from confirmed HSV-1 genital infections (first episodes and recurrent infections). Detection of HSV-2 antibody in 21 sera from HSV-2 first episodes was more effective by WBA which identified homologous antibody in 19 (96%), whereas MAb block RIA detected HSV-2 antibody in 16 (76%). HSV-2 antibody was detected in 24/25 (96%) sera from recurrent HSV-2 infections by WBA and by MAb block RIA, the highest degree of concordance for both methods. In addition, the MAb block RIA may be more effective in detecting the presence of HSV-1 antibody in sera from recurrent HSV-2 cases. Prevalence of HSV-1 and HSV-2 antibody was measured by the MAb block RIAs in 3 UK human study populations which consisted of 100 children/young adolescents, 104 adult blood donors and 80 genito-urinary medicine clinic attenders.

Adolescent↗
PubMed1995

Automated microneutralization: method and comparison with western blot for type-specific detection of herpes simplex antibodies in two pregnant populations.

A novel method for the detection of microneutralization was compared to Western blot for detection of type-specific antibodies to herpes simplex virus in two pregnant patient populations in Vancouver. From an unselected group of women in labor in a tertiary care obstetric hospital (426) and another group of women specially referred for assessment of genital herpes in pregnancy (195) 20 and 88%, respectively, were seropositive for herpes simplex virus type 2 antibodies by Western blot. During the study period, 93 (48%) of the high-risk group were culture positive for HSV-2 and all but one case was verified serologically by Western blot. Therefore, the sensitivity for Western blot in this high seroprevalence group was 99%. The false-negative case had longstanding recurrent infection and on retest was found to be a laboratory error. For microneutralization, type-specificity was especially difficult to distinguish among patients with mixed viral type patterns. Comparing microneutralization to Western blot as the gold standard, sensitivity, specificity, positive and negative predictive values were 84, 78, 97 and 40% respectively, for the referred group and 73, 82, 51 and 92%, respectively, for the unselected group. Poor negative and positive predictive values for the referred group and unselected populations, respectively, demonstrate that microneutralizations are not clinically acceptable for type-specific antibody detection. By contrast, serology by Western blot is very sensitive and apparently highly specific.

Antibodies, Viral↗
PubMed1994

Protein-specific cervical antibody responses to primary genital herpes simplex virus type 2 infections.

Antibodies to herpes simplex virus (HSV) have been demonstrated in cervicovaginal secretions but have not been analyzed for their viral protein targets, prevalence, isotype, or kinetics of development. A method was devised to collect cervical secretions from women with primary genital HSV-2. By Western blot, cervical IgG and IgA responses to HSV-2 proteins VP5, gB, and gD were detected in most patients within 2 weeks of onset and to gC/gE within 3 weeks. Cervical IgM and IgG responses to gG, VP16, and ICP35 developed later and were more variable. Cervical IgM to most proteins appeared within 6-10 days. Cervical IgA and IgG persisted for weeks, but cervical IgM waned. Western blot profiles of serum IgG and cervical IgG to individual HSV-2 proteins were similar; those of serum IgA and cervical IgA differed. These findings suggest a universal, complex immune response to HSV-2 infection in the female genital tract.

Adult↗
PubMed1993

Presumptive identification of common adenovirus serotypes by the development of differential cytopathic effects in the human lung carcinoma (A549) cell culture.

The neutralization test is commonly used in clinical virology laboratories for the identification by serotype of adenovirus isolates. In an effort to conserve reagents and reduce the amount of time in the performance of this assay, we evaluated the significance of differential cytopathic effects for the presumptive identification of lower-numbered adenovirus serotypes that are commonly encountered in the clinical setting. Utilizing the human lung carcinoma (A549) cell culture as our indicator system, two viral induced monolayer degenerations (i.e., cytopathic effects or CPEs) were recognized. Among our wild and the laboratory adapted (i.e., ATCC) adenovirus isolates tested in this study, serotypes 1, 2, 4, 5, 6, 8, 11, 19, 21, 27, and 31 were expectedly characterized by the typically enlarged, rounded, and refractile cells, which eventually aggregated into irregular 'grape-like' clusters. Adenovirus types 3 and 7, however, were characterized by the development of distinct intranuclear inclusions, a flattening and then a web or net-like monolayer degeneration. Differences in the intensity of intranuclear granulation were related by electron microscopy to differences in the quantity of viral crystalline aggregates within the host cell nucleus. A presumptive identification of the commonly encountered adenovirus serotypes 3 and 7 prior to the performance of the neutralization test would result in a conservation of type-specific antiserum, a decreased use of cell cultures and medium, and lastly, reduced medical technologist workload.

Adenoviruses, Human↗