摘要:

AbstractThe expression of nine oncogenes (c‐myc, N‐myc, N‐ras, H‐ras, k‐ras, abl, fos, src, and raf) and two tumor suppressor genes (p53 and RB) were studied by northern blot hybridization in six human hepatocellular carcinoma or hepatoblastoma cell lines (PLC/PRF/5, HepSB, Hep G2, 2.2.15, HLE, and HLF) and in a human embryonic lung fibroblast cell line (WI‐38) to look for differences that might be associated with the presence (PLC/PRF/5, Hep3B, and 2.2.15) or absence (Hep G2, HLE, and HLF) of integrated hepatitis B virus (HBV) DNA. The levels of expression of the oncogenes and tumor suppressor genes were unrelated to the presence or absence of integrated HBV‐DNA. Furthermore, the intensity of expression of these oncogenes was no greater in the 2.2.15 cell line (consisting of Hep G2 cells transfected with hepatitis B virus) than in untransfected Hep G2 cells. © 1992 W

ISSN:0146-6615    

DOI:10.1002/jmv.1890380402

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractNewly available HBV serological assays have not been established routinely in most underdeveloped countries. Utilizing enzyme‐immune assays to determine the presence of pre‐S1 antigen and anti‐pre‐S2, and using two conventional hybridization techniques and the PCR assay to detect HBV‐DNA, we studied 30 HBsAg chronic carriers and as a reference group 10 subjects whose only HBV routine marker was anti‐HBc. Seventynine percent of the HBeAg positive carriers showed detectable HBV‐DNA by a non‐radioactive slot‐blotting technique. The PCR assay was more sensitive than the slot‐blotting technique, detecting HBV‐DNA in anti‐HBe positive patients with moderate or normal ALT activity. Pre‐S1 antigen was mostly related t o the presence of HBsAg and anti‐pre‐S2 was associated with active viremic state, increased ALT activity (ranges 51 to 640 IU/L), and with self‐limited HBV infection. The presence of HBV‐DNA i n the group with anti‐HBc only was detectable solely by the PCR assay. For an underdeveloped country the addition of a PCR assay or pre‐S1 anti‐pre‐S protein tests to the current assessment procedures of HBV chronic infection should be used only i n selective cases. HBeAg/anti‐HBe serological evaluation and HBV‐DNA detection by a non‐isotopic conventional hybridization technique still remain as useful tools to screen initially for the presence of viremia in chronic HBsAg carriers. The presence of HBV‐DNA in individuals with anti‐HBc only suggests that anti‐HBc screening should be maintained and expanded to all the blood banks of less industrialized countries where the rate of HBV infection in apparently he

ISSN:0146-6615    

DOI:10.1002/jmv.1890380403

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractA modified centrifugation culture technique and a polymerase chain reaction (PCR) is described for detection of early antigen and IE antigen DNA, respectively, for rapid and sensitive monitoring of active cytomegalovirus (HCMV) infection after organ transplantation.In a preliminary study, 541 clinical specimens (blood, urine, bronchoalveolar lavage, pharyngeal wash, sputum) from 59 organ recipients were assayed for HCMV antigen by centrifugation culture; 144 samples were tested by PCR simultaneously. Antigenemia detected by centrifugation culture correlated strongly with active HCMV infection and clinical symptoms and proved useful for monitoring the efficacy of anti‐viral therapy. PCR was more sensitive in an earlier phase of infection when centrifugation culture was still negative. The clinical usefulness of both methods is discussed. © 1992 Wiley‐Liss,

ISSN:0146-6615    

DOI:10.1002/jmv.1890380404

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractIn two studies comparing detection of human cytomegalovirus (HCMV) in 118 patients (93 of whom were immunocompromised) by the polymerase chain reaction (PCR) and virus isolation using either early antigen detection by culture‐immunofluorescence or conventional cytopathic effect, DNA‐PCR was found to be the most sensitive, followed by culture‐immunof luorescence, then by cytopathic effect. Urine was inhibitory to the action of Taq polymerase; this was overcome by concentration of HCMV with PEG 6000 prior to gene amplification. Without PEG treatment, HCMV‐DNA in 6 of the 11 specimens positive by culture‐immunofluorescence was not detectable by PCR. In healthy seropositive individuals, HCMV‐DNA was not detected i n leucocytes. However, in immunocompromised patients with AIDS or transplants, and therefore at high risk of HCMV infection or reactivation, blood leucocytes were usually positive for HCMV‐DNA (19/20), some for as long as 20 weeks after initial detection and persisting for long after culture‐immunofluorescence became negative. Neither HCMV‐RNA nor infectious HCMV were detected in the follow‐up blood leucocycte specimens from immunocompromised patients who had detectable HCMV‐DNA in these cells. These data suggest that persistence of HCMV‐DNA in blood leucocytes of immunocom‐promised patients after reactivation or primary infection may be due to persistence of non‐viable virus, residual HCMV genomic DNA, or latent HCMV

ISSN:0146-6615    

DOI:10.1002/jmv.1890380405

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractThe ability of varicella zoster virus (VZV) to infect and replicate in human keratinocytes in culture was examined. Primary human keratinocytes derived from the abdomen, breast, and foreskin were plated as monolayers and infected by co‐cultivation with VZV infected fibroblasts (MRC‐5 cells). Replication and spread of the virus was assayed by plaque assay and immunofluorescence of infected cells using a VZV specific monoclonal antibody. Although all three types of keratinocytes tested were capable of supporting productive VZV infection, the keratinocytes showed a 1.5 to 2 log reduction in virus yield as compared to infection of monolayer cultures of MRC‐5 cells. Results from immunofluorescence studies and plaque assays indicate a slower rate of cell‐to‐cell spread of the virus. Testing of an anti‐VZV compound in this novel assay system demonstrated an interesting sensitivity compared to that observed in conventional assay Systems. © 1992 Wil

ISSN:0146-6615    

DOI:10.1002/jmv.1890380406

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractThe use of urine as a noninvasive specimen for the diagnosis of hepatitis A (HAV) and hepatitis B (HBV) virus infections was investigated. Specimens of urine were collected at the same time as blood or saliva specimens, or singly in cases of previously serologically confirmed recent infection. The specimens were tested for IgG and IgM anti‐HAV and anti‐HBc by immunoglobulin class‐specific capture radioimmunoassays (GACRIA and MACRIA). On the basis of assays on urine specimens it was possible to distinguish between individuals who were susceptible or immune to HAV or who had recently been infected with HAV. Using assays on 327 corresponding saliva specimens as reference tests, the observed sensitivity and specificity of tests on urine specimens by anti‐HAV GACRIA were 98.9% and 99.1%, respectively, and by anti‐HAV MACRIA were 95.8% and 99.6%, respectively. IgM and IgG anti‐HBc were detected readily in the urine of 35 acute or recent cases of hepatitis B but were not found in the urine of seronegative individuals. Of the urine specimens from 52 individuals who were HBsAg carriers or who had had long past HBV infections, 49 contained detectable IgG anti‐HBc. Of urine specimens from 42 HBsAg carriers, 11 contained raised IgM anti‐HBc levels.Urine, which is a convenient specimen to collect, can be used t o study outbreaks of hepatitis A, to ascertain the HAV immune status of individuals, to differentiate hepatitis A from hepatitis B, and to identify individuals who have been naturally exposed to HBV. © 1992

ISSN:0146-6615    

DOI:10.1002/jmv.1890380407

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractAstroviruses are intestinal pathogens associated with gastroenteritis in man and animals. The mechanism of internalization into host cells has not been reported previously. The cell entry pathway of serotype 1 human astrovirus into 293 cell line was studied biochemically and morphologically.Viral infection was monitored by indirect immunofluorescence. Infected cells were treated with the lysosomotropic agents ammonium chloride, methylamine, and dansylcadaverine or the ionophore monensin to raise the intraendosomal and intralysosomal pH. All drugs tested inhibited the early stages of infection whereas they did not interfere with the viral binding to the plasma membrane.The presence of astrovirus particles was detected by electron microscopy in coated pits and later in coated vesicles.The data indicate adsorptive endocytosis as the most probable mechanism by which astroviruses enter susceptible cells. © 1992 Wiley‐Liss, I

ISSN:0146-6615    

DOI:10.1002/jmv.1890380408

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractVirological and serological studies were carried out prospectively to evaluate the possible activation of human herpesvirus‐6 (HHV‐6) in 50 infants and children with acute measles by isolation of HHV‐6 from peripheral blood and by determining neutralizing antibodies to the virus. All but 5 patients (90%) were seropositive to HHV‐6 in the acute stage of measles and 18 (40%) had a significant increase in HHV‐6 antibody titers thereafter, whereas only 2 of 27 patients who were initially seropositive to Epstein‐Barr virus (EBV) viral capsid antigen (VCA) had a significant rise in antibody titers to EBV VCA. Among 18 patients with a significant increase in HHV‐6 titers, the virus was isolated from the peripheral blood mononuclear cells of three patients in the early convalescent stage of measles. These results indicate that activation of HHV‐6 may occur frequently a few weeks after primary infection with the measles virus. © 1992

ISSN:0146-6615    

DOI:10.1002/jmv.1890380409

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractDirect detection of respiratory syncytial (RS) virus antigen i n nasopharyngeal secretions (NPS) provides the most rapid diagnostic test for RS infection, but more sensitive methods might be more beneficial in the study of virus shedding. RS virus RNA was extracted from cells stored at −70°C either in suspension with added RNAse inhibitor or as a pellet without inhibitor. The RNA was reverse transcribed, the resultant cDNA amplified by the polymerase chain reaction and detected by ethidium bromide staining after electrophoresis through agarose gel (RT‐PCR). Of 217 specimens tested, 106 were positive by antigen detection, 99 by RT‐PCR, and 92 by virus isolation. In a series of 97 sequential NPS specimens from 15 infants in whom RS virus induced bronchiolitis was confirmed, antigen detection proved most sensitive in the first week after on‐set and RT‐PCR detected most positive specimens in the second week. Storage of the cells as a pellet proved more satisfactory than storage as a suspension. A further round of amplification using nested primers increased the number of positive results obtained by RT‐PCR. The sensitivity of antigen detection using directly labelled monoclonal antibody to RS virus was slightly higher than that of RT‐PCR, but the specificity was slightly lower. © 1992

ISSN:0146-6615    

DOI:10.1002/jmv.1890380410

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY

摘要:

AbstractThe association of hepatitis C virus (HCV) infection and tattooing was studied in 87 tattooed and 126 tattoo free healthy young men who did not engage in intravenous drug use or multiple sexual activity. Antibody against HCV (anti‐HCV) was tested in serum specimens by enzyme immunoassay with C100‐3, NS3, and core antigens; 11 of the 87 (12.6%) tattooed and 3 of the 126 (2.4%) tattoo free subjects were positive for anti‐HCV (odds ratio = 5.9, 95% CI = 1.6–22.0). A relationship was demonstrated by an increased risk for HCV infection with an increasing number of tattooed site (Ptrend= 0.002). All but one of the 87 tattooed subjects had been infected by hepatitis B virus (HBV) and 25 were carriers of hepatitis B surface antigen (HBsAg). None of the 25 HBsAg carriers was positive for anti‐HCV whereas 11 of the 62 HBsAg non‐carriers had anti‐HCV, suggesting a negative association between the HBsAg carriage and the long lasting anti‐HCV (P= 0.02, Fisher's exact). The status of the tattooer was also an important determinant for HCV infection; the risk was higher if tattooing was done by a non‐professional friend than by a professional tattooist. Tattooing, probably with improperly sterilized needles, can clearly pose an increased risk for HCV infection in Taiwan. This study indicates the need for legal standards for hygienic tattooing as part of preventive measures for the control of parenterally transmitted infections. © 19

ISSN:0146-6615    

DOI:10.1002/jmv.1890380411

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1992

数据来源: WILEY