Viral studies of mothers with human immunodeficiency virus infection at delivery and their infants in the first 3 days of life.

OBJECTIVE: We studied 49 mother-infant pairs for human immunodeficiency virus (HIV) (a) to assess the virological and immunological status of HIV-infected mothers at delivery and their infants within the first 3 days of the infant's life, and (b) to correlate these findings with eventual infection outcome in the infant. METHOD: Maternal blood from women in labor and infant's blood within 3 days of life were tested for the titer of HIV immunoglobulin G (IgG) antibody, for presence of HIV by culture, for p24 antigen, for HIV DNA by polymerase chain reaction (PCR), and for absolute T-helper cell count (CD4). RESULTS: Eight infants were in the confirmed infected (CI) group, with a transmission rate of 21%. Thirty infants were in the confirmed uninfected (CU) group. In the mother, mean anti-HIV IgG titer was 1:2600 (CI group) and 1:3350 (CU group); in the infant, the mean titer was 1:3250 (CI group) and 1:2710 (CU group). Eighty-seven percent of the mothers were culture-positive in the CI group compared to 33% in the CU group (p = 0.005). Eighty-seven percent of CI infants were PCR-positive at birth; none was PCR-positive in the CU group (sensitivity = 87%; specificity = 100%). Sixty-two percent of CI infants were culture-positive at birth, whereas none was positive in the CU group (sensitivity = 62%; specificity = 100%). Of the uninfected infants, 23% were positive for p24 antigen at birth. CONCLUSIONS: HIV IgG antibody titers in mothers and their infants at birth were markedly elevated in both CI and CU groups but were not protective against infection. However, the high titers explain the long duration of this antibody in the blood of infants born to infected mothers. Culture positivity in the mother at delivery correlated highly with eventual infection in the infant (p = 0.005). HIV antigen, specifically p24 antigen, was detectable in uninfected infants when tested at birth.

Perinatal 'TORCH' infections identified by serology: correlation with abnormalities in the children through 7 years of age.

A matched case-control methodology was used to assess the risk for a wide range of abnormalities in children associated with serological evidence for 'TORCH' infections in the mothers. Specimens were selected from the large bank of sera from the approximately 54,000 pregnant women who participated in the Collaborative Perinatal Project. There was no clear association between any of the antigens studied and any specific damage to the child. These 'negative' findings are consistent with the absence of frequent significant effects due to these agents in the second and third trimesters of pregnancy.

Toxoplasmosis: maternal and pediatric findings in 23,000 pregnancies.

An analysis of the antibody titers to toxoplasmosis for 22,845 pregnant women in the Collaborative Perinatal Project was conducted in relation to clinical and laboratory findings in the mothers and children through 7 years of age. More than 900 observations were considered for each mother and child. The major findings were in the children and included a predicted doubling in the frequency of deafness among children born to women with antibody to toxoplasmosis, a predicted 60% increase in microcephaly, and a 30% increase in low IQ (less than 70) in association with the presence of high maternal antibody titer (256 to 512) to toxoplasma. A serologically defined high-risk group of mothers was identified on the basis of high indirect hemagglutination antibody levels or seroconversions and increased IgM toxoplasma antibody levels (indirect fluorescent antibody greater than or equal to 32, enzyme-linked immunosorbent assay greater than or equal to 0.7). Of the 15 pregnancies in this group, two children had congenital toxoplasmosis and three were stillborn.

Avidin-biotin latex agglutination assay for detection of antibodies to viral antigens.

A new method for attaching antigens to latex by an avidin-biotin technique is described. The procedure permits control of the amount of antigen attached to the latex and eliminates the need for highly purified antigens and destructive bridging chemicals. The avidin-biotin latex agglutination assay is a simple, rapid test well suited to detection of viral antibody. The sensitivity and specificity, respectively, of the avidin-biotin latex agglutination assay compared with other assay results for antibody to viruses were as follows: cytomegalovirus, 98 and 100% (indirect hemagglutination assay); measles virus, 96 and 100% (enzyme-linked immunosorbent assay); and herpes simplex virus, 78 and 100% (indirect hemagglutination assay).

Serologic studies of MS patients, controls, and patients with other neurologic diseases: antibodies to HTLV-I, II, III.

We have studied the frequency of human retrovirus antibody (HTLV-I, II, III) in the serum and CSF of patients with MS, matched controls, and patients with optic neuritis, idiopathic and postencephalitic Parkinson's disease, neuropathies, polymyositis, ALS, and postpoliomyelitis. Except for the postpoliomyelitis samples, all samples were collected prior to 1980. Contrary to a previous published report, no significant levels of antibody to HTLV-I, II, or III were found in the MS patients or controls. No retrovirus antibody was detected in patients with the other neurologic diseases.

Antibody to human and simian retrovirus, HTLV-I, HTLV-II, HIV, STLV-III, and SRV-I not increased in patients with multiple sclerosis.

We have tested sera from patients with multiple sclerosis, matched controls, and those with other neurological diseases, as well as sera from patients with the acquired immunodeficiency syndrome and controls and patients with tropical spastic paraparesis (TSP) and controls for antibody to human T-lymphotropic virus type I (HTLV-I), HTLV-II, human immunodeficiency virus (HIV), simian T-lymphotropic virus type III, or simian retrovirus type I by immunofluorescent activity test, and for HTLV-I and HIV by the ELISA method. Sera from patients with multiple sclerosis and matched controls, and from patients with optic neuritis and Parkinson's or other neuromuscular diseases did not have antibody to any of the retroviruses tested. Specimens from TSP patients and some controls contained HTLV-I antibody. We conclude from our study that only TSP patients had serological evidence of infection with one of the retroviruses studied.

HIV and HTLV-I antibody studies: pregnant women in the 1960s, patients with AIDS, homosexuals, and individuals with tropical spastic paraparesis.

To investigate the possible occurrence of human immunodeficiency virus (HIV) or human T-cell lymphotropic virus, type I (HTLV-I) infections in the United States prior to 1979-1981, when acquired immune deficiency syndrome (AIDS) was first recognized, we tested sera from 310 pregnant women who participated in the Collaborative Perinatal Project during the period 1959-1964 for HIV and HTLV-I antibody. These samples included sera from 53 pregnant women who were intravenous drug users. The remainder were from women who had cervical epithelial abnormalities, who developed cervical carcinomas, who had had children with erythroblastosis fetalis, who had had children that developed malignant neoplasms early in life, or normal pregnant women. None of the 310 women had confirmed HIV or HTLV-I antibody. The rate of false-positive reactions with the HIV enzyme-linked immunosorbent assay (ELISA) antibody test in these long-frozen samples was similar to that observed in fresh sera. HIV antibody was detected in homosexual patients with AIDS; HTLV-I antibody was not detected in any of these sera. HTLV-I antibody was detected in 17 of 20 patients with tropical spastic paraparesis (TSP) and in two of seven patients with other neurological diseases diagnosed as transverse myelopathy and multiple sclerosis, and in none of nine normal controls; HIV antibody was not detected in any of these sera patients. Thus, we conclude that there was no serological evidence of infection with HIV or HTLV-I in the pregnant women studied; however, HIV antibody was present in all AIDS patients tested, and HTLV-I antibody was found in the majority of patients with TSP.

Capillary enzyme immunoassay for rapid detection of herpes simplex virus in clinical specimens.

Capillary enzyme immunosorbent assay (CapELISA) is a modification of the standard enzyme immunosorbent assay which permits rapid detection of viral antigens in clinical specimens. The capillary tube format provides a very large reactive surface relative to the sample size. The close proximity of antigen to antibody in the tube optimizes the reaction, resulting in increased sensitivity and shorter incubation requirements. Sensitivity is further enhanced by use of a biotin-avidin enzyme detector system and a fluorogenic rather than a colorigenic substrate. The assay is performed at ambient temperature and requires less than 2 h. It is read with an inexpensive hand-held black light. Data for the use of CapELISA for the detection of herpes simplex virus in clinical specimens are presented. The results show greater than or equal to 85% sensitivity and 100% specificity when compared with tissue culture tests on the same samples. This new system should be advantageous for the diagnosis, treatment, and management of patients with herpesvirus infections and for the prevention of neonatal herpes acquired by passage of the fetus through an infected birth canal.

Difficulties associated with serological diagnosis of Toxoplasma gondii infections.

Physicians often rely on serology to help determine whether a patient has had a recent infection with Toxoplasma gondii and as an aid in estimating the possible teratogenic effect on the fetus. For this reason the diagnostic laboratory should take every precaution to avoid misleading results. The best serological analysis is based on a rise in IgG titer with two appropriately spaced serum samples. Also, the presence of a high IgM titer in one serum sample is generally considered to be good evidence that infection has occurred recently. The indirect fluorescent antibody (IFA) test has been the most widely used test for detection of IgG or IgM. Recently enzyme-linked immunosorbent assays (ELISA) have also been developed for this purpose. In this study we reaffirm that false IgM positive results can occur with these tests because of the presence of rheumatoid factor in serum, and false negative results can also occur because of competitive inhibition by specific IgG. We show that a preabsorption of serum with a Staphylococcus/Streptococcus preparation (Staffinoc, MA Bioproducts, Walkersville, MD) removes IgG and IgA and eliminates many of the false reactions. We have also found that elevated levels of specific IgM can persist for at least several years in some women. This suggests that the presence of IgM alone is not always an indication of recent infection.

Immunoglobulin M antibodies detected by enzyme-linked immunosorbent assay and radioimmunoassay in the diagnosis of cytomegalovirus infections in pregnant women and newborn infants.

Immunoglobulin M (IgM) antibodies were detected by a commercially available enzyme-Linked Immunosorbent Assay (ELISA) in 36 of 49 (73%) pregnant women with primary cytomegalovirus (CMV) infection. A positive ELISA-IgM result occurred in 10 of 13 patients (77%) assessed within 8 weeks of seroconversion. The sensitivity of the radioimmunoassay (RIA) to identify primary infection in pregnant women was comparable, 78% in general and 86% for women tested within 16 weeks of seroconversion. Of the 36 women with primary infection who had detectable IgM antibodies by ELISA, 25 (69%) were delivered of congenitally infected infants, whereas of the 13 with undetectable IgM antibodies, 7 (54%) transmitted the infection in utero. IgM antibodies were detected by ELISA in only 5 of 43 (11%) women who experienced a recurrence of CMV which either did or did not result in congenital infection. RIA was less likely to measure CMV-specific IgM in recurrent infection, inasmuch as 1 of 19 (5.2%) women with proven recurrent infection had detectable IgM antibody, giving RIA a better specificity for primary infection. Specific IgM antibodies were detected by ELISA in 42 of 61 (69%) babies congenitally infected with CMV and in 4 of 70 (5.7%) uninfected control newborn infants. The RIA was superior for diagnosis of congenital CMV infection, with a sensitivity of 89% and a specificity of 100%. The lower sensitivity of the ELISA-IgM occurred in the category of congenitally infected infants born to mothers with recurrent infection (43%), a group that is at the lowest risk of disease or to develop sequelae. This commercially available ELISA-IgM could be used in combination with a CMV-specific IgG test for monitoring women during pregnancy for primary infection.

Enzyme-linked immunosorbent assay for detection of measles antibody.

An enzyme-linked immunosorbent assay (ELISA) was developed for the detection of measles immunoglobulin G antibody (MEASELISA). This assay was found to be comparable to the measles hemagglutination inhibition (HAI) test. Approximately 500 sera from three centers were tested by MEASELISA and the HAI test. MEASELISA demonstrated values of greater than 99% for sensitivity, specificity, and accuracy. Values were very precise, with a mean coefficient of variation of 5.4%. MEASELISA values were shown by linear regression analysis to increase as HAI titers increased. A coefficient of determination of 1.00 was obtained from test center three. MEASELISA values were found to be linearly related (r2 greater than 0.97) to MEASELISA titers, thus enabling quantitation of measles antibody from a single value. Also, data are presented that show MEASELISA to be equivalent to complement fixation for evaluating paired sera for the presence of a significant increase in antibody levels to measles virus.

Antibody-dependent cell-mediated cytotoxicity against Epstein-Barr virus membrane antigen in African Burkitt's lymphoma.

Serial samples of sera from patients with African Burkitt's lymphoma were tested for antibody against Epstein-Barr virus (EBV)-specific membrane antigen (MA) by the antibody-dependent cell-mediated cytotoxicity assay (ADCC). Titers of patients in the long-term survivor group were generally higher than those found in the sera of patients in the short-term survivor group. Although ADCC titers to EBV-MA were not useful in predicting which patients would relapse there was a definite relationship between ADCC titer and prognosis. The individual differences in ADCC titers in patients in remission may explain the variability of responses that have been reported in studies on serotherapy with remission plasma.

Enzyme immunoassay for detection of antibody to Epstein-Barr virus-specific early antigen.

Antibody to Epstein-Barr virus-induced early antigen could be measured in sera using enzyme-linked antibody. The sensitivity of this assay was comparable to that of the indirect immunofluorescent technique. Measurement of early antigen was accomplished on a producer cell line which was specifically treated to block late gene expression. It is now possible to measure Epstein-Barr virus-induced early antigen and viral capsid antigen antibodies by using the same cell line.

Comparison of indirect hemagglutination and indirect immunofluorescence tests with microneutralization tests for detection of type-specific Herpesvirus hominis antibody.

Indirect hemagglutinating and immunofluorescent antibody responses to Herpesvirus hominis types 1 and 2 were compared to neutralizing antibody responses in infected humans from whom H. hominis type 1 or 2 was isolated. The indirect immunofluorescent antibody test was shown to be the most sensitive and specific for primary human infections. The sensitivity and specificity of the indirect hemagglutination and the immunofluorescent antibody tests were shown to be equal to that of the microneutralization test among patients who had primary or recurrent H. hominis type 2 infections. It is suggested that the indirect hemagglutination test is preferable for assaying large populations for previous infection with H. hominis type 2 because it is rapid, easier to perform, and more economical. The intermediate range of titer differences (deltat) between H. hominis types 1 and 2 previously reported to be due to infections with both viruses was shown to occur in all three tests among patients with primary infections with either virus.

Simultaneous radioimmunoassay for specific antibodies to members of the human herpesvirus group.

A method of described for the simultaneous radioimmunoassay (RIA) for antibody to members of the human herpesvirus group. The RIA is compared with some of the conventional serologic techniques used to quantitate antibody to these viruses (Epstein-Barr virus, cytomegalovirus, herpesvirus type 1 and varicella-zoster virus). Color-coded beads, each coated with the antigens of a different herpesvirus, were similtaneously placed in a well which contained a human serum to be assayed for antibody to each of these 4 viruses. The results of this test were compared with the results obtained when the serum was assayed for antibody to the 4 viruses in 4 separate tests. We conclude that the antigen-antibody reactions do not significantly interfere with each other when a serum is assayed for antibody to the 4 viruses simultaneously. A comparison of the RIA with conventional serologic techniques shows excellent correlation in the antibody titers obtained. Features of the solid-phase RIA allow significant savings of time, reagents and space, and thus make it feasible for the small laboratory to screen large numbers of sera for antibody to a variety of antigens.

Brain tumors in owl monkeys inoculated with a human polyomavirus (JC virus).

Owl monkeys were inoculated intracerebrally, subcutaneously, and intravenously with JC, BK, or SV40 virus. Two of four adult owl monkeys inoculated with JC virus, a human polyomavirus, developed brain tumors at 16 and 25 months after inoculation, respectively. A grade 3 to grade 4 astrocytoma (resembling a human glioblastoma multiforme) was found in the left cerebral hemisphere and brainstem of one monkey. The second monkey developed a malignant tumor in the left cerebral hemisphere containing both glial and neuronal cell types. Impression smears prepared from unfixed tissue of this tumor showed cells that contained polyomavirus T antigen. Virion antigens were not detected. Tumor cells cultured in vitro also contained T antigen but were negative for virion antigen. Infectious virus was not isolated from extracts of this tumor.

Antigenic relationship of two strains of simian hemorrhagic fever virus.

Serological comparison of the prototype and an epizootic (Corbell) strain of simian hemorrhagic fever virus revealed that the two viruses were serologically similar. The prototype strain differs from the Corbell strain in that the latter cannot be cultivated in vitro. Serological comparison of the prototype virus grown in tissue culture and its homologous antibody and the prototype and Corbell viruses recovered from rhesus monkey serum and their homologous antibodies showed differences and suggest that a complex relationship exists which has not yet been defined.

The etiology of non-specific urethritis in active duty Marines.

We studied patients with non-specific urethritis and control subjects at our dispensary. These patients and controls were matched for age, rank and sexual activity, and studied for the presence of bacteria, virus, Trichomonas and Mycoplasma. No significant bacteria were found in either group. No Trichomonas was identified, only 1 herpes II was recovered and no cytomegalovirus was found. Mycoplasma were cultured from patients and controls. The rate of colonization varied, depending upon several factors, but the significant factors seemed to be associated with the number of sexual partners. Those men with more than 1 sexual partner had a significantly increased colonization with Mycoplasma.