Progression of prevalence of HIV-1 infection among injection drug users in Montreal, Quebec.
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Biomedical subjects
Publications and source records attributed to M Fauvel.
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We have used the polymerase chain reaction (PCR) to detect HIV proviral sequences in minute amounts of peripheral blood collected onto newborn screening blotters. Forty-three newborns, infants, and children of HIV-infected mothers were serially studied: dried blood spot (DBS) specimens were processed for PCR; serum was assayed for HIV antibodies, p24 antigen, and immunoglobulins; mononuclear cells were cultured and CD4 cells were quantitated by immunofluorescence. There was excellent agreement between the results of blood spot PCR, viral culture, and clinical and immunological indicators of HIV infection. Eighteen of 19 infected children tested positive by both PCR and culture, including six asymptomatic infants who were less than 10 weeks of age. As expected, p24 antigen capture assays were insensitive, detecting only 13 of the 19 infected children. One infected infant tested positive by PCR, but negative by culture and antigen. This infant was seropositive at 27 months and had pronounced hypergammaglobulinemia in association with non-specific symptoms. Twenty-four of the 43 infants were asymptomatic with normal immune profiles, declining antibody levels and no evidence of infection. These children tested repeatedly negative by PCR, culture, and p24 antigen assays. Our results indicate that DBS PCR is a sensitive, specific, and cost-effective alternative to viral culture for the early diagnosis (or exclusion) of perinatal HIV infection. DBS sampling opens the way for large-scale prospective studies to determine the exact rates of vertical HIV transmission in industrialized, as well as, nonindustrialized countries.
Endocaval filters are often used to prevent pulmonary embolism but they have a number of disadvantages. The DIL filter, made of a memory metal wire, is intended to male up for some of these disadvantages. It acts by modifying the shape of the inferior vena cava, which it filters through its meshed loops. It is inserted percutaneously, causes little trauma, and its release is progressive. However, it requires measuring the caliber of the inferior vena cava. This filter was inserted in thirty-four patients over a period of 13 months. One filter has migrated. No recurrence of pulmonary embolism and no thrombosis of the inferior vena cave occurred.
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Electron microscopy and an enzyme-linked immunosorbent assay were compared for the diagnosis of rotaviruses associated with neonatal calf diarrhea. One hundred percent correlation was observed when 125 samples were tested by both techniques. Both techniques were equally efficient in detecting rotaviruses. The enzyme-linked immunosorbent assay was more sensitive but being specific could not detect other viruses.
Rotaviruses have been shown to be of importance as aetiologic agents of gastroenteritis in infants and in domestic animals of several species. Hemagglutinins were prepared from two Canadian isolates of bovine rotavirus and from one isolate of a simian rotavirus. A United Kingdon isolate of bovine rotavirus was shown not to possess hemagglutinating activity, indicating a strain difference between a Canadian and United Kingdom bovine rotavirus. In hemagglutination-inhibition (HAI) tests a rabbit hyperimmune (two injections) serum, prepared to one of the bovine rotaviruses, was not helpful in distinguishing the two bovine viruses because of cross-reactions between the viruses. However, it was possible to distinguish the bovine viruses from the simian virus with this serum. When guinea pig immune sera were prepared to the four rotavirus strains and tested with the three hemagglutinins in the HAI test, antigenic differences between the four strains of rotavirus were demonstrated. Hyperimmune guinea pig serum prepared to a strain of human rotavirus did not inhibit any of three hemagglutinins indicating that the human strain is different from the three rotavirus strains which gave hemagglutinins.
A hemagglutinin has been prepared from Nebraska calf diarrhea virus (NCDV) propagated in BS-C-1 cell line. After cesium chloride centrifugation, the hemagglutinin of the bovine rotavirus was found to be associated with intact virions (density 1.355 g/ml) but not with virions lacking an outer capsid layer (density 1.375 g/ml). In hemagglutination-inhibition (HAI) tests, the hemagglutinin reacted specifically with NCDV serum, and HAI seroconversions were detected in some sera tested. Cross-reactions were observed in complement fixation tests between the human and bovine rotaviruses but were not demonstrated by HAI, suggesting that the hemagglutinin detects a specific rather than a group antibody response.
Counterimmunoelectrophoresis was compared with electron microscopy for detection of human reovirus-like agent in fecal specimens. Both tests gave very similar results.
Rotaviruses are generally difficult to isolate and culture in vitro; therefore, virus isolation has not been used as a method of diagnosing this group of agents. The present report describes a simple procedure for isolating bovine rotaviruses directly from feces after pretreatment of fecal samples with trypsin. This procedure resulted in virus isolation from five of five samples that contained virus particles, as demonstrated by electron microscopy, and four of seven samples where virus particles could not be observed but were considered positive by the presence of immunofluorescent-staining cells in feces. Virus could not be isolated from "normal" feces. If the virus was not passaged in the presence of trypsin, the infectivity was gradually lost, but infectivity could be restored again if trypsin was added, resulting in increased virus spread and concomitant increase in virus yield. The application of this technique as a diagnostic tool for bovine and other rotaviruses is briefly discussed.
Representative members of four arbovirus taxons were studied by immune electron microscopy (IEM) in an attempt to determine whether the technique can be applied to the specific identification of arboviruses. Alphavirus, flavivirus, bunyavirus, and rhabdovirus antigens were prepared by propagation in tissue culture. The viruses were used to titrate homologous antibody and test heterologous antibody in the IEM test. Specific clumping together with visualization of antibody attachment was observed for all homologous systems studied. For the alphaviruses, titers of antibody, as determined by IEM, were comparable with the titers obtained by hemagglutination inhibition. Alphavirus cross-reactions were observed at low antibody dilutions by IEM. However, further dilution of antibody resulted in specific clumping only of homologous virus, allowing specific identification of alphavirus unknowns. For all viruses studied, positive IEM reactions were obtained with homologous grouping ascitic fluids but not with heterologous grouping fluids.
Hepatitis B core antigen (HBcAg) is found on the decoated Dane particle and on a morphologically similar particle detected mainly in the nucleus of hepatocytes of patients with hepatitis B. HBcAg prepared from the liver of a chimpanzee infected with hepatitis B virus was used to test human serum for core antibody (anti-HBc) by complement fixation. Anti-HBc was found in serum collected from patients with hepatitis B in both the acute and convalescent stages, from carriers of hepatitis B surface antigen (HBsAg) and from patients with chronic liver or renal disease who were carriers of HBsAg. It was not found in patients with hepatitis A or infectious mononucleosis, or in healthy persons who were not carriers of HBsAg.
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Hepatitis B surface antigen (HBsAg) positive plasma, obtained from blood donors, was used to prepare an antigen (HBcAg) from the inner core of the Dane particle, the larger of the two spherical particles found in the blood in hepatitis B infection. The antigen was prepared by a series of centrifugation procedures on caesium chloride and sucrose gradients. Dane particle-rich fractions obtained from these gradients and concentrated by centrifugation were treated with Tween 80 to remove the outer coats from the particles. The remaining Dane particle cores were recovered by velocity sedimentation on a sucrose gradient. Preparations containing the Dane cores were used as antigens in the immune electron microscopy and complement fixation tests to demonstrate the specificity of HBcAg and lack of fross relationship with HBsAg. HBcAg will be used for the study of hepatitis B infections in man.