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Biomedical subjects

J V Parry

Publications and source records attributed to J V Parry.

At least 55 records · Page 3Linked to original sources

Performance of the Amplicor human immunodeficiency virus type 1 PCR and analysis of specimens with false-negative results.

Over a 4-year period, the Roche Amplicor kit was used in a United Kingdom reference laboratory for the detection or confirmation of human immunodeficiency virus (HIV) type 1 infection, particularly in infants born to HIV-infected mothers. Of 408 specimens from adults and older children tested, the 122 seronegative specimens were all Amplicor negative. Of the 286 seropositive specimens, 268 were Amplicor positive. On the basis of these results, the Amplicor assay has a specificity of 100% and a sensitivity of 93.7%. In addition, for 247 specimens from infants and young children, serological results may not have been diagnostic because of placental transfer of maternal antibodies. Forty-eight were Amplicor positive, and of the 199 Amplicor-negative specimens, 19 were assumed to be false negative on the basis of clinical data, serological markers (including p24 antigen), and/or results for previous or follow-up specimens. This represents a sensitivity of 75% for the Amplicor test for specimens from patients under 2 years of age. Of these 37 false-negative specimens plus 2 specimens from other laboratories, 31 could be characterized by amplifying extracted material from them by an in-house nested gag PCR spanning the Amplicor target region. The amplicons were sequenced and found to represent subtypes A (35.5%), B (22.6%), C (22.6%), D (16.1%), and G (3.2%). False-negative results by the Amplicor assay may be ascribed to low-target copy number, the physical behavior of one primer (SK462), and sequence variation in the target region of the other primer (SK431).

Adult↗

Continuing transmission of sexually transmitted diseases among patients infected with HIV-1 attending genitourinary medicine clinics in England and Wales.

OBJECTIVE: To determine whether those who are aware of being infected with HIV continue to adopt behaviours that place others at risk of HIV infection. DESIGN: Ongoing survey of current diagnosis of sexually transmitted disease and awareness of HIV infection among patients attending genitourinary medicine clinics. SETTING: Six genitourinary medicine clinics in England and Wales (two in London and four outside) participating in unlinked anonymous HIV serosurveillance during 1990-3. SUBJECTS: All attenders having blood drawn for syphilis serology for the first time during the calendar quarter of attendance. MAIN OUTCOME MEASURES: The proportion of syphilis serology specimens with antibody to HIV-1 detected by unlinked anonymous testing of the residue. The proportion of attenders infected with HIV-1 who remained clinically undetected, and the proportion who had another recently acquired sexually transmitted disease. RESULTS: Of 85441 specimens tested, 2328 (2.7%) were positive for antibodies to HIV-1. About 30% of these specimens were from attenders whose HIV-1 infection remained clinically undetected. HIV-1 infection was found to coexist with another recently acquired sexually transmitted disease in 651 attenders, of whom 522 were homosexual or bisexual men. Of these, 245 (47%) already knew themselves to be infected with HIV-1. This proportion increased between 1990 and 1993. CONCLUSIONS: A considerable proportion of patients infected with HIV-1 are not identified by voluntary confidential HIV testing in genitourinary medicine clinics. Substantial numbers of homosexual or bisexual men attending genitourinary medicine clinics continue to practise unsafe sex despite being aware of their infection with HIV-1.

Acquired Immunodeficiency Syndrome↗

How valuable are IgA and IgM anti-HIV tests for the diagnosis of mother-child transmission of HIV in an African setting?

BACKGROUND: Babies born to HIV-infected mothers retain anti-HIV of maternal origin until 15-18 months of age. Because of this, HIV proviral DNA and p24 antigen measurements have become the methods of choice for timely diagnosis of HIV infection in infancy. They are, however, too expensive for widespread use in the developing world. OBJECTIVE: To evaluate a simple, inexpensive serological method for diagnosing mother-child transmission of HIV, in an African population, which takes account of the effects of placental transfer of maternal antibody and continued exposure to HIV through breast-feeding. STUDY DESIGN: Plasma specimens for a prospective study of mother-to-infant transmission of HIV in rural Zaire were collected at birth, 3, 6, 9, 12, 18 and 24 months from 21 infected infants (PP group), 21 uninfected infants (PN group) born to seropositive mothers and 21 control infants (NN group) born to uninfected mothers. The specimens were retrospectively tested for IgG, IgM and IgA anti-HIV by immunoglobulin class-specific capture EIAs, and by a commercial anti-HIV EIA. RESULTS: In neonatal specimens, IgA and IgM anti-HIV were present, respectively, in 13 of 14 (97%) and 8 of 14 (57%) of the PP group and in 6 of 11 (55%) and 2 of 11 (18%) of the PN group. Later, at 3 months and older, IgA and IgM anti-HIV were only detected in the PP group. They peaked at 18 months (93%) and 24 months (67%) respectively. Of the 21 PP group children, 8 (38%) were transiently IgG anti-HIV-negative in the first year, indicating that infection had probably taken place after birth; four of the 8 had no detectable IgA anti-HIV during the first year. None of the specimens collected from the NN group babies were reactive for IgA, IgM or IgG anti-HIV. CONCLUSIONS: IgA and IgM anti-HIV may be passively transferred across the placenta. Where breast-feeding is prevalent, about half of the transmissions may occur after birth, thus delaying the diagnosis of mother-child transmission. Nevertheless, this simple, cheap IgA anti-HIV, EIA identified 65% of transmissions by 9 months of age, and 93% at 18 months of age. It is a more useful marker than IgM anti-HIV, and gave a much more rapid answer than did tests for IgG anti-HIV seroreversion.

Journal Article↗

HIV-1 prevalence in community-wide samples of injecting drug users in London, 1990-1993.

OBJECTIVE: To provide time-trend estimates of HIV-1 prevalence among injecting drug users (IDU) in London. DESIGN: HIV-1 prevalence and HIV testing behaviour were measured in four serial point prevalence surveys of IDU recruited at multiple sites in community-based non-treatment and drug treatment settings between 1990 and 1993. METHODS: Community subjects were recruited through social network sampling by trained indigenous interviewers; treatment subjects were interviewed at agencies. With informed consent, subjects responded to a structured questionnaire covering risk behaviour. Volunteered saliva samples were tested anonymously for anti-HIV-1. Statistical comparisons across years were examined using mixed binomial logistic and log-linear models. Pearson's chi 2 and Fisher's exact tests were also used for some two-group comparisons. RESULTS: Similar samples were recruited each year. HIV-1 prevalence rate declined from 12.8% in 1990, 9.8% in 1991, 7.0% in 1992, to 6.9% in 1993. The statistical modelling suggested that the overall trend in prevalence rates was one of decelerating decline. There was no difference in prevalence rate by gender and length of injecting. Less than one-half (46%) had received a named HIV test. Over one-half of the HIV-positive IDU were unaware of their status. CONCLUSIONS: The pattern of decline in HIV prevalence rate is attributed to changes in risk behaviour following HIV prevention interventions.

Adult↗

Salivary testing for human immunodeficiency virus type 1 infection in children born to infected mothers in Sao Paulo, Brazil. The Sao Paulo Collaborative Study for Vertical Transmission of HIV-1.

OBJECTIVE: To validate a method for salivary testing for HIV infection in children older than 12 months. METHODS: Oral fluid samples were collected via sponge foam swabs from children born to HIV-positive mothers and were tested for antibodies to HIV-1 and HIV-2 with an IgG antibody capture enzyme-linked immunosorbent assay and a modified Western blot for confirmation. In each child serum antibody status was the standard used to validate the salivary antibody test. RESULTS: We obtained 331 oral fluid samples from children born to HIV-positive mothers. The specificity and sensitivity of salivary testing compared with results on sera were both 100% (297 of 297 (95% confidence interval 98.8 to 100%) and 34 of 34 (95% confidence interval 89.7 to 100%), respectively). Compliance in the study population increased from 91% to 97% when mothers were offered the opportunity to provide oral fluid from their children instead of blood specimens. CONCLUSION: Salivary testing provides an accurate and acceptable noninvasive method for assessing the HIV infection status of children born to infected mothers by using IgG antibody capture enzyme-linked immunosorbent assay alone with a strategy of duplicate retesting of reactive specimens.

Acquired Immunodeficiency Syndrome↗

Variation in the sensitivity of HBsAg screening kits.

Fifteen HBsAg kits from 14 manufacturers were assessed. Their sensitivity was evaluated by testing 150 HBsAg-positive sera, sera from four donors who were low-level HBsAg carriers, and sequential specimens from 22 seroconverting individuals together with dilutions of six of these specimens. The British HBsAg Working Standard (0.5 IU mL-1) and the NIBSC/UKBTS HBsAg Monitor Sample (0.125 IU mL-1) were also tested. Five assays failed to detect one of the 150 routine HBsAg-positive sera. Four assays (Auszyme Monoclonal; Monolisa Ag HBs 2nd generation; Murex HBsAg; Ortho HBsAg Test Systems 3) were able to detect HBsAg in all but one of the six sera from low-level carriers, whereas one assay (MicroTrak II HBsAg) detected only one of the six. The most sensitive kit (Monolisa Ag HBs 2nd generation) detected HBsAg in 79 specimens from the seroconversion panels; four other kits detected HBsAg in at least 70 specimens, seven in 60-69, two in 50-59 and the least sensitive in 31. Further analysis of the findings on seroconverters indicated a median reduction in the duration of HBsAg detection of 5 days or more for four assays when compared with the most sensitive assay. One kit (Auszyme Monoclonal) detected HBsAg in 15 of the 18 dilutions prepared from the seroconversion specimens, whereas three kits detected HBsAg in fewer than 10 dilutions. Two kits gave negative reactions with the British HBsAg Working Standard on all of five occasions and six were consistently unreactive with the NIBSC/UKBTS HBsAg Monitor Sample; only three kits (Bioelisa, Enzygnost, Murex) were always reactive. There is therefore substantial variation in sensitivity among the HBsAg kits currently available.

Evaluation Studies as Topic↗

Anti-HIV antibody in saliva: an assessment of the role of the components of saliva, testing methodologies and collection systems.

The various components of saliva, namely mixed saliva, parotid saliva, submandibular saliva, crevicular fluid and minor (labial) gland secretions, were collected from 63 known HIV antibody seropositive patients. A commercial test system, Wellcozyme HIV 1+2, and an antibody capture ELISA (GACELISA), were compared for sensitivity against all components. Sensitivity of the GACELISA system was 100% in 123 mixed saliva, 121 parotid saliva and 127 labial fluid samples, and 98% in 99 submandibular samples and 127 crevicular fluid samples. Respective figures for Wellcozyme 1+2 were 92%, 55%, 73%, 66% and 63%. Mixed saliva was most easily, conveniently and effectively collected using a plain Salivette. In 241 Salivette samples examined from the 63 patients, GACELISA proved 100% sensitive, and Wellcozyme 95% sensitive. Another form of Salivette impregnated with citric acid was unsuitable for GACELISA and gave a false negative value of 45%. In 197 samples from the gingival margin taken by a dry swab, GACELISA showed a sensitivity of 98% and Wellcozyme 81%. The most sensitive method for demonstrating anti-HIV antibody in saliva is to collect mixed saliva with the plain Salivette system and assay anti-HIV antibody levels by GACELISA.

Adult↗

The rising prevalence of HIV-1 infection in patients attending an inner city accident and emergency department.

The recently published findings of the unlinked anonymous HIV prevalence study in England and Wales showed unchanging HIV prevalence in groups such as homo/bisexual men, and declining rates in non-injecting heterosexual men attending genitourinary medicine clinics. However, this multicentre study did detect a significant rise in seroprevalence rates in pregnant women in England and Wales and sentinel groups within hospitals in London, warning that changing patterns of HIV infection might account for these variable results. In 1992-1993 a seroprevalence study of adult patients attending the accident and emergency department at St. Mary's Hospital in West Central London showed a rate of HIV-1 infection of 1 in 77. We have repeated the seroprevalence study over the same calendar months in 1994-1995 to gain further information about HIV positive patients attending the department and to see whether a change in the patterns of HIV infection in the population served by St Mary's Hospital had occurred.

Adult↗

A collaborative case control study of sporadic hepatitis A in England.

A case control study of sporadic hepatitis A was carried out in 201 districts in England from July 1990 to June 1991. The aims were to determine the risk factors associated with the infection and to identify individuals or groups who might benefit from prophylactic measures, such as human normal immunoglobulin or hepatitis A vaccine. Factors associated with risk of hepatitis A included travel abroad (odds ratio (OR) 19.8; 95% confidence interval (CI) 4.87-80.6), a household contact with hepatitis A (OR 13.5; 95% CI 6.49-28.0) and sharing a household with a child aged 3 to 10 years (OR 1.57; 95% CI 1.1-2.22). This study provided no clear evidence of increased risk in health care workers, teachers, or other occupational groups. A non-significant trend towards an increased risk in nursery nurses and child minders aged 20 to 29 years was observed. Pre-exposure prophylaxis with hepatitis A vaccine may be considered for people who travel frequently to areas where hepatitis A is highly or moderately endemic. Post-exposure prophylaxis with human normal immunoglobulin should be given to contacts of known cases in accordance with national guidelines. Immunoglobulin alone has been recommended in outbreaks, but the use of vaccine alone or combined with immunoglobulin should be evaluated.

Adolescent↗

The risk of hepatitis A from sewage contamination of a water supply.

The drinking water supply of a town became contaminated with sewage. The town's population was surveyed to determine the baseline prevalence of hepatitis A antibodies and to establish whether an associated outbreak of hepatitis A had occurred. Samples of saliva were obtained from 540 people in 200 randomly selected households, and tested for IgG and IgM antibodies to hepatitis A. Fifty-six per cent (279/495) were susceptible to hepatitis A and 43% (213/495) immune as a result of previous infection. Immunity was directly related to age; those who were immune were significantly older than those who were susceptible (mean ages: 43.5 years and 19.0 years; p < 0.0001). Six people were found to have had a recent infection with hepatitis A, but exposure to hepatitis A during the water pollution incident was possible in only one case. The results offer no evidence that this incident caused an outbreak of hepatitis A, but the study has provided useful epidemiological data on hepatitis A.

Adolescent↗

GACPAT HIV 1 + 2: a simple, inexpensive assay to screen for, and discriminate between, anti-HIV 1 and anti-HIV 2.

A simple and cheap assay suitable for screening for anti-HIV 1 and anti-HIV 2 and discriminating between them was evaluated. In it specimens are incubated in U-bottomed microplate wells coated with anti-human IgG for 30 min at room temperature. After washing, 100 microliters of a 1 in 50 dilution of HIV 1-coated gelatin particles (Serodia-HIV 1/2, Fujirebio) are added. Settling patterns are read on the second day: A positive reaction is indicated by adherence of the particles and a negative by a button. The HIV 1 particles are then washed away and HIV 2 particles added. Anti-HIV 2 reaction patterns are read on the third day. To assess the performance of the modified "GACPAT HIV 1 + 2" assay a panel of 1,621 serum/plasma specimens was used. It comprised validated anti-HIV 1 positive (n = 220), anti-HIV 2 positive (n = 214), dual anti-HIV 1/anti-HIV 2 positive (n = 11), and anti-HIV negative (n = 1,176) serum/plasma specimens. All 434 specimens that contained anti-HIV 1 or anti-HIV 2 reacted positively with the homologous particles. The 11 dually positive specimens reacted positively with both HIV 1 and HIV 2 particles. Five (2.3%) anti-HIV 1 and five (2.3%) anti-HIV 2 positive specimens gave positive reactions with both particle types, but none of the five cross-reactive anti-HIV 2 specimens were dually reactive when the order of particle addition was reversed.(ABSTRACT TRUNCATED AT 250 WORDS)

AIDS Serodiagnosis↗

Investigation of a hepatitis A outbreak in a primary school by sequential saliva sampling.

BACKGROUND: An outbreak of hepatitis A occurred in a primary school (children aged 4-11 years), starting in the Autumn of 1990 and terminating some 5 months later after some spread into the local community. OBJECTIVES: The objectives were to monitor the spread of the virus within the primary school over time, to document infection in asymptomatic individuals and the efficacy of using saliva for HAV antibody detection in young children as an acceptable screening method by using the Salivette method and ordinary cotton tipped swabs. STUDY DESIGN: Serial saliva samples were taken over a period of months and anti-HAV IgM and IgG antibodies measured by radioimmunoassay. RESULTS: Twenty-seven children in the school and nine individuals from the surrounding community acquired hepatitis A. Twenty-one (78%) of the 27 children were symptomatic, as were all the affected adults. The cotton-tipped swabs were found to be as effective a method as Salivette at diagnosing infection in those in whom the methods were compared. CONCLUSIONS: Despite many reports stating that children are more likely to be asymptomatic with HAV infection we found the majority to report significant symptoms. Young children do not easily accept serum sampling as a method for diagnosis or epidemiological studies, and we show that saliva sampling is an effective and acceptable diagnostic method.

Journal Article↗

Testing for antibody to human immunodeficiency virus type 1 in a population in which mycobacterial diseases are endemic.

During a large epidemiologic study in the Karonga District of northern Malawi, serum samples from 139 patients with incident leprosy, 124 with newly diagnosed leprosy, 277 patients with incident tuberculosis, and 2296 controls were tested for antibodies to human immunodeficiency virus. Sera were tested according to a four-test protocol using two ELISAs and two particle agglutination assays. Overall, 188 samples were considered positive, 2634 were considered negative, and 14 were indeterminate. All 18 available positive specimens from leprosy patients, a random sample of 14 positive specimens from tuberculosis patients, and 15 positive specimens from controls were tested by Western blot. There was no evidence of substantial numbers of ELISA false-positives in any patient group or among controls.

Adult↗

Evaluation of a combined HIV-1/2 and HTLV-I/II assay for screening blood donors.

A combined immunoassay for the simultaneous detection of antibodies to HIV-1/2 and HTLV-I/II (Bioelisa, Launch Diagnostics) has been evaluated to determine its suitability for routine use in blood screening. 84,222 donations were tested from 76,452 donors. One HIV- and 1 HTLV-1-positive donor were identified. The specificity was 99.7%, and the sensitivity for anti-HIV-1, anti-HIV-2, and anti-HTLV-1 on 173 positive sera was 100%; 2 of 25 anti-HTLV-II-positive sera were non-reactive. Although the specificity of the assay is not as high as that of HIV-1/2 kits currently in UK transfusion use, the information gained about donor HTLV antibody status makes the test an attractive alternative to them.

Blood Donors↗

False negative results in enzyme linked immunosorbent assays using synthetic HIV antigens.

The sensitivity of 12 commonly used anti-HIV-1/HIV-2 enzyme immunoassays was evaluated. The assays, each of which utilises at least one synthetic HIV antigen, were tested against a panel of 1092 specimens previously designated anti-HIV positive. In a total of 13 104 tests there were eight false negative results attributable to assay insensitivity: three were on two serum samples collected close to seroconverison and five on another serum specimen. These eight false negative results arose in seven different assays. Five other false results were attributable to technical error. This false negativity rate indicates that all of the assays performed adequately and leads to an estimate of one false negative result in a thousand tests in routine diagnostic practice. Because of the antigenic heterogeneity of HIV strains, similar evaluations would be required in several regions before this satisfactory level of sensitivity in anti-HIV assays incorporating synthetic antigens could be said to be universal.

Enzyme-Linked Immunosorbent Assay↗

Management of blood donors whose donations are repeatedly falsely positive by the HIV antibody screening test.

Since 1985, over 1,800,000 donations have been screened by the West Midlands Regional Blood Transfusion Service for antibody to HIV. Twelve regular donors gave three or more donations that were alternatingly positive and negative in the screening test, but not confirmed to be HIV positive by supplementary testing. Extensive investigation of six of these donors, including the polymerase chain reaction (PCR), failed to confirm HIV infection. The donors were reassured but, nevertheless, retired to comply with the guidelines of the National Blood Transfusion Service. These findings indicate that, for UK donors, ambiguous serological findings are unlikely to reflect HIV infection. On the rare occasions where serological results are particularly ambiguous, PCR testing of donors' blood may be helpful.

AIDS Serodiagnosis↗