[Role of cytomegaloviruses (CMV) in blood transfusions--virologic and immunologic aspects].
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Antibodies, Viral↗
Biomedical subjects
H W Doerr
Publications and source records attributed to H W Doerr.
[Possibilities in the virological laboratory diagnosis of CNS infections].
Viral infections of the central nervous system (CNS) are investigated by the use of a great lot of laboratory methods preferring organ-specific patients' material, f. e. cerebrospinal fluid (CSF). Beside of the improvement of virus isolation procedures, antigen tests and assays on nucleic acid will get more importance for rapid viral diagnosis. Pathognomonic intrathecal antibody formation can be demonstrated by immunoglobulin class and subclass differentiation in CSF specimens. However, some of the slow virus diseases must still be retrospectively analysed by the means of a neuropathological laboratory.
Antibodies, Viral↗
A whole-blood lymphoproliferation assay for measuring cellular immunity against herpes viruses.
A whole blood test system was established to study cell-mediated immunity to cytomegalovirus (CMV) and herpes simplex virus (HSV) in a large number of healthy blood donors. Cellular immunity was measured by the in vitro proliferative response (LP) of peripheral lymphocytes. These responded vigorously to several mitogens. Lymphocytes of most individuals responded to HSV, but only a limited number were reactive towards CMV. In parallel, antibodies against CMV and HSV were measured by an ELISA technique. For HSV, good correlation was observed between serological and lymphocyte proliferation results. For CMV, no clear correlation was obtained, only 21 of 40 donors positive in the antibody test being positive in the LP test. The majority of seronegatives were negative in the LP test. Use of virions purified by sucrose gradient centrifugation, or an additional strain of CMV (strain Davis) did not increase the number of donors positive in the LP test. One explanation might be that individuals possessing antibodies against CMV as measured by ELISA but no capacity to react in the LP test had suffered from a CMV infection a long time before, and now showed waning cellular immunity, but antibody still detectable. Use of the whole blood technique on 108 individuals showed that this very simple test works well with various mitogens and at least some antigens.
Antigens, Viral↗
Human cytomegalovirus infection: recent developments in diagnosis and epidemiology.
Cytomegalic inclusion disease (CID) is caused by a horizontally or vertically transmitted human herpes virus infection and may persist for life without obvious clinical symptoms. A serious course of horizontal primary and recurrent infections, however, is often observed in immunocompromised persons such as recipients of organ transplants and patients receiving fresh blood transfusions. Vertical infection may cause fetopathies. The human cytomegalovirus (HCMV) is thought to inherit an oncogenic potential as lately discussed for AIDS and M. Kaposi. Laboratory diagnosis of HCMV infection is performed by light microscopy (inclusion bodies), electron microscopy, virus isolation in cell culture, demonstration of viral DNA and antigen in clinical specimens, by histochemical methods (e.g. immunoperoxidase technique) and by DNA and peptide analysis for identification of different isolates and viral finger prints. Evaluation of cell-mediated immunity in HCMV infection is performed quantitatively (assessment of Thelper/Tsuppressor ratios) or qualitatively (specific lymphocyte stimulation by the antigen). In most cases laboratory diagnosis is achieved by serological methods, i.e. demonstration and quantitation of HCMV-specific antibodies. In this context, a number of liquid- and solid-phase immunoassays have been developed, of which immunofluorescence and ELISA are most commonly used, besides complement fixation and passive haemagglutination. These procedures on the one hand allow the use of different antigen preparations as early and late viral proteins, and on the other hand permit a specific determination of different Ig classes and subclasses. A variety of assays has been established especially for determination of virus-specific IgM antibodies, which are predominantly found in active infection. These, however, at least in part may show non-specific results caused by interference of rheumatoid factor or IgG competition. Such problems have now been dealt with and are avoided by IgG precipitation or IgM immunosorption ("mu-capture" technique). These recent methods allow an exact epidemiological identification of risk groups for CMV infection. Results from our laboratory revealed 13% HCMV-IgM positive patients among pregnant women, 16% IgM positive patients among renal transplant recipients, 4% IgM positive cases in patients after cardiosurgery and 1.7% IgM positives among prostitutes. The prevalence of HCMV infection as indicated by specific IgG antibodies was 56%, 90%, 83%, and 90%, respectively.(ABSTRACT TRUNCATED AT 400 WORDS)
Acquired Immunodeficiency Syndrome↗
Detection of human rotavirus by nucleic acid analysis in comparison to enzyme-linked immunoassay and electron microscopy.
The results of RNA analysis for the detection of rotavirus were compared with those of a standard enzyme-linked immunosorbent assay (ELISA) and electron microscopy using 212 faecal specimens obtained from 200 children with gastroenteritis. Rotavirus was extracted directly from faecal specimens and RNA segments were made visible by polyacrylamide gel electrophoresis using a silver staining technique. Of the 212 faecal specimens 137 were found to be positive in ELISA, 125 in RNA analysis and 121 in both methods. Forty-nine of the 212 specimens were also investigated by electron microscopy. Thirty-five were positive when examined by electron microscopy, 37 were positive in ELISA and 33 in RNA analysis. RNA analysis of 119 faecal samples in outbreaks and sporadic cases of rotavirus infection yielded 42 different rotavirus electrophoretypes. The results indicated that no one method was sufficient to detect all positive specimens and that RNA analysis is useful in epidemiological studies.
Electrophoresis, Polyacrylamide Gel↗
Incidences of antibodies to hepatitis B, herpes simplex and cytomegalovirus in prostitutes.
The 170 prostitutes registered by the local public health authority (165 female, 5 male; ages 17-61) were serologically investigated by ELISA and CFT for the presence of hepatitis B virus (HBV) markers (n = 170), anti-CMV (n = 118) and anti-HSV 2 antibodies (n = 117). 4.7% (1.2%) were found to be HBs- (HBe-)antigen carriers; 1.2% revealed an acute hepatitis B (seroconversion of anti-HBc or IgM-anti-HBc highly positive); 17.1% presented a past and resolved hepatitis B (anti-HBs positive). These rates exceeded those of 125 female age-matched control persons considerably, out of whom only 2.4% (0.8%) presented HBs-(HBe-) antigen and 3.2% anti-HBs antibodies. 90% of the prostitutes investigated proved to be anti-CMV IgG (1.7% IgM) antibody positive and were compared to healthy blood donors (47% IgG, 0% IgM-anti-CMV antibody carriers) and other CMV risk groups: pregnant women 56 (13), patients in hemodialysis 61 (0), hemophiliac patients 69 (0), patients after kidney transplantation 90 (16) and after open-heart surgery 87 (4) % anti-CMV IgG (IgM) antibody carriers. An AIDS patient suffered from simultaneous HBV and CMV infections. The risk of HSV 2 infection is among prostitutes (38.5% seropositives) five times as high as among female age-matched control persons (7.2% seropositives, n = 125). These results confirm that prostitutes must be regarded as a risk for the spread of hepatitis BV and CMV- and HSV 2-caused diseases in the population.
Adolescent↗
Cytomegalovirus (CMV) infections in renal transplant recipients. Preliminary results of prophylaxis by an intramuscular human hyperimmune CMV IgG.
Infectious diseases are the most serious complications in immunosuppressed patients and the major cause of death in renal transplant recipients (23, 44). Besides bacterial, fungal and protozoan infections, viruses of the human herpes group (herpes simplex virus, varicella-zoster virus, Epstein-Barr virus, cytomegalovirus) are known to be of major importance (21, 37). The most common and clinically relevant virus of this group is the cytomegalovirus (CMV).
Adolescent↗
Determination of HBV DNA by a simplified method of spot hybridization.
Molecular hybridization was employed to detect HBV DNA in sera of patients with acute or chronic hepatitis, by a simplified version of the spot hybridization technique. HBV DNA was found in 21 out of 50 sera obtained in acute hepatitis B. Determination of HBV DNA was negative in sera of patients with hepatitis A, Epstein-Bar virus infections or other HBsAg-negative liver diseases. There was no cross-hybridization between HBV DNA and sera of patients with non-A, non-B hepatitis.
Acute Disease↗
[Anti-HBc IgM in acute and chronic hepatitis B virus infection].
Hepatitis B core antigen (HBcAg) synthesized in E. coli was used for determination of immunoglobulin M class-specific antibodies against HBcAg. It was found that 98% of cases with acute hepatitis B surface antigen (HBsAg) positive hepatitis type B were anti-HBc immunoglobulin M (IgM) positive. Atypical hepatitis B was detected in 33% of anti-HBc-positive HBsAg-negative cases with acute hepatitis. Anti-HBc IgM was positive for 6 months in acute resolving hepatitis type B, whereas cases resulting in chronic hepatitis B remained anti-HBc IgM-positive for up to 900 days. Chronic HBsAg carriers with severe liver disease had anti-HBc IgM more often than individuals with minor liver damage; 83% of HBsAg-positive liver cirrhoses, 63% of chronic aggressive hepatitis, 50% of HBsAg-positive liver carcinoma, but only 17% of chronic persistent hepatitis or 7% of healthy blood donors were anti-HBc IgM-positive. Determination of anti-HBc IgM is useful in detecting atypical hepatitis B virus infections without HBsAg in serum and, with some restrictions, in discriminating acute and chronic hepatitis type B.
Acute Disease↗
Use of hepatitis B core antigen produced in Escherichia coli to detect immunoglobulin M specific antibodies in an enzyme-linked immunosorbent assay.
The antigenic activity of HBcAg produced in Escherichia coli and HBcAg from human liver was compared in a mu-specific solid-phase antibody-capture assay for detection of anti-HBc-IgM. HBcAg from liver could be detected in dilutions up to 1:3, HBcAg from Escherichia coli in dilutions up to 1:10,000. Using HBcAg from Escherichia coli, sera from five patients with acute resolving hepatitis B and sera from four patients with acute hepatitis B who had developed chronic liver disease were tested for anti-HBc-IgM in ELISA. IgM fractions separated out of the same sera by immunoaffinity chromatography were tested for anti-HBc-IgM using a commercially available test. The results were in good agreement with those obtained by ELISA. Anti-HBc-IgM could be detected up to 900 days after onset of disease. Different groups of patients were tested for presence of anti-HBc-IgM in ELISA. Fifty-nine of 60 patients with acute hepatitis B were positive for anti-HBc-IgM at onset of illness. Ten of 16 patients with chronic aggressive hepatitis and seven of 23 HBsAg positive dialysis patients were also positive for anti-HBc-IgM, whereas only two of 12 patients with chronic persistent hepatitis and one of 15 HBsAg positive blood donors ("healthy" carriers of HBsAg) had detectable anti-HBc-IgM.
Enzyme-Linked Immunosorbent Assay↗
HBV-DNA in sera of patients with HBsAg-positive primary liver cell carcinoma.
Sera of ten patients with HBsAg-positive primary liver carcinoma were tested for anti-HBc-IgM and HBV-DNA. Five patients were positive for anti-HBc-IgM and six for HBV-DNA. There was no correlation between the presence of anti-HBc-IgM and HBV-DNA. Our study suggests that complete viral replication exists in some HBsAg-positive primary liver carcinomas.
Aged↗
[Demonstration of Rubella-specific IgG and IgM antibody (sub) classes using the enzyme immunoassay (EIA)].
The sensitivity of three different enzyme-linked immunosorbent assays (ELISAs: Enzygnost-Rubella/Behring Co., Rubazyme (M)/Abbott Co., Rubelisa M/M.A.B.) was examined for detection of rubella-specific IgM and IgG antibodies. The screening of the sera of 20 s.c. vaccinated persons (rubella strain Cendehill) using these diagnostic systems revealed that the ELISAs are capable to detect the rubella IgG antibodies reliably. Additional haemagglutination inhibition (HI), and single radial haemolysis (SRH) tests confirmed this fact. Antibody kinetics seen in Enzygnost-Rubella are similar to those determined by HI. In contrast, it was found that the Rubazyme test is able to detect largely late IgG antibodies in agreement with the data obtained by SRH, which may be helpful to exclude a rubella reinfection. A result negative in Rubazyme, but positive in HI should indicate a positive rubella IgM test. For the determination of subclass-specific IgG antibodies the sandwich assay was extended to a 4-phases-test, using specific antibodies to IgG1-IgG4. After acute rubella infection or vaccination the antibodies of the IgG3 subclass appear before the other. The rubella antibodies in the IgG3 exceed its quantitative distribution in the total IgG (about 7.5%). The determination of IgM-specific rubella antibodies with the Enzygnost-Rubella and Rubazyme M systems proved also to be rather sensitive and reliable. No interference was seen from rheumatoid factor activity, IgG competition and heterologous IgM. In contrast, the Rubelisa M system should be improved in evaluation procedures. However, since all systems showed also differences to HI using IgM fractions, it seems to be advisable to apply an additional rubella IgM test beside ELISA in critical cases.
Adult↗
[Rubella, measles, mumps. Possibilities of rapid diagnosis].
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Adolescent↗
Rapid fractionation of serum immunoglobulins by high pressure liquid gel permeation chromatography. Application to routine serologic procedures.
High pressure liquid chromatography (HPLC) was applied to separation of IgM from IgG and IgA for the detection of virus-specific antibodies by routine serologic methods. Serum samples of 250 microliter were fractionated on a HPLC protein column after filtration for use in rubella haemagglutination inhibition after preabsorption with kaolin. The protein fractions were examined for IgM, IgG and IgA content and for cross contamination. The relative recovery after kaolin absorption was satisfactory: IgM greater than 60%, IgG greater than 90%, IgA greater than 90%. The immunoglobulin M was well separated. IgG/IgA contamination of the fraction was less than 0.2%/ less than 0.5%. The IgM and IgG fractions were used without further treatment in haemagglutination inhibition, complement fixation and enzyme-linked immunosorbent assay. Highly specific results in the diagnosis of acute viral diseases (Rubella, Herpes zoster, cytomegalovirus) and Mycoplasma pneumoniae were obtained.
Antibodies, Bacterial↗
[Rubella infection in pregnancy. Methods of determining risk].
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Adolescent↗
Non-A, non-B hepatitis in hemodialysis.
Episodes of presumable non-A, non-B hepatitis were determined among hemodialysis patients during a two year period as well as possible secondary infections in household contacts. We determined hepatitis A and B markers by commercial RIA kits, and IgM-anti-CMV and IgM-anti-EBV by ELISA techniques. 154 dialysis patients, 118 relatives of patients, and 42 members of the staff were included in the study. 71% of the center dialysis patients, 63% of the home dialysis patients, 50% of the staff, and 19% of the relatives were HBV marker positive. Spouses of patients more often had HBV markers than other relatives. Anti-HAV was highly prevalent both in center patients (98.8%) and in home dialysis patients (83.3%). The incidence of IgM-anti-CMV was marked in home dialysis patients (15.5%). During a two year period, 9% of all dialysis patients had an episode of presumable non-A, non-B hepatitis. As compared to previous data in the same dialysis centers, there is a change in the epidemiology of hepatitis.
Adult↗
Follow-up of needlesticks in medical staff.
In an on-going study, 104 events of needlestick or contamination of apparent skin lesions were evaluated for the risk of hepatitis. Whenever possible, a blood sample of the 'donor' was screened for HBsAg as well as for elevated transaminases. Members of the staff were bled immediately after exposure and were followed up for 9 months. At present the follow-up is completed in twenty probands. - Hepatitis B immunoglobulin (= HBIG) was given in 51 cases. HBsAg was detected in 21 donors and strongly supposed in 27 cases. Only one nurse developed acute hepatitis B ten months after exposure to HBsAg-positive blood. 53 probands were exposed to HBsAg-negative blood. 32 persons received SIG. Only 5 'donors' were classified as probable Non-A, Non-B hepatitis which did not induce apparent infections in passively immunized persons. 23 persons did receive neither HBIG nor SIG. 2 persons had been exposed to HBsAg-positive blood by needlestick, still being without seroconversion. Passive immunization after needlestick should be performed after testing blood samples of donors and recipient. After exposure to non-A, non-B hepatitis, secondary infections did not appear in passively immunized persons.
Female↗
Hepatitis markers in a psychiatric institution.
Hepatitis A and B markers were determined in 714 patients and in 291 members of the staff of a psychiatric institution for adults. The leading diagnosis were psychosis (71.8%), neurosis (17.7%), and oligophrenia (10.6%). Anti-HAV was found in 84.4% of the patients and in 69.2% of the staff. 26.8% of the patients and 19.9% of the staff had at least one HBV marker. The frequency of HBV markers correlated with the age of the patients but not with the duration of the hospitalization. After the exclusion of IgM-anti-HAV, IgM-anti-CMV and IgM-anti-EBV or possible toxic effects, 2.6% of all patients remained as presumable non-A, non-B hepatitis infections. In this study HBV markers were not found in an hyperendemic pattern reported previously in psychiatric institutions.
Adult↗