urine exposure to hbv
Sep 1, 2010
i have been reading in the BRITISH MEDICAL JOURNAL about urine exposure to hbv the discussion was about 23 men with chronic liver disease who were positive for hepatitis B antigen and they are saying that it is possible to be infected from there urine
below is the clinical research article , i would appreciate if you could read this and ripply to me with your opinion, thanks!
Hepatitis B virus DNA in saliva, urine, and seminal fluid of carriers of hepatitis B e antigen
Concentrated samples of saliva, urine, and seminal fluid from 23 men with chronic liver disease who were positive for hepatitis B e antigen were examined for the presence of hepatitis B virus deoxyribonucleic acid (HBV-DNA) by molecular hybridisation. HBV-DNA was detected in saliva from 15 of 17 men (88%), urine from 12 of 22 men (55%), and seminal fluid from 13 of 21 men (62%). The presence of hepatitis B virus in such secretions has important epidemiological implications for heterosexual and homosexual contact. Introduction The parenteral mode of transmission of hepatitis B virus is well established, but non-parenteral (or inapparent parenteral) routes of exposure may also play a part in infectivity; this may explain the high incidence of transmission of hepatitis B virus among homosexual men and family contacts of carriers of hepatitis B virus.'-7 Several groups have therefore examined bodily fluids and secretions for the presence of hepatitis B surface antigen (HBsAg), which has been found in saliva,8-10 urine,'-"2 semen,8 breast milk,13 14 vaginal secretions and menstrual blood,15 16 and pancreatic and biliary secretions."7 There are conflicting reports of its presence in faeces.10 18-20 The detection of HBsAg alone, however, does not by itself denote potential infectivity as HBsAg may be found in the absence of infectious virions. The potential for transmission of Academic Departments of Medicine and Physiology, Royal Free Hospital School of Medicine, London NW3 2PF P KARAYIANNIS, Bsc, PHD, research assistant D M NOVICK, MD, research fellow A S F LOK, MB, MRCP, research fellow M J F FOWLER, Bsc, PrD, research assistant J MONJARDINO, MB, Prm, reader in physiology H C THOMAS, PrD, FRCP, professor of medicine Correspondence to: Professor H C Thomas. hepatitis B virus by inapparent parenteral means has therefore not been fully evaluated. The presence of hepatitis B virus particles can be detected by molecular hybridisation techniques, which detect hepatitis virus deoxyribonucleic acid (HBV-DNA).2"-25 The presence of HBV-DNA indicates potential infectivity. In this study we examined concentrated saliva, urine, and seminal fluid for the presence of HBV-DNA by molecular hybridisation. The results were correlated with serum concentrations of HBV-DNA and with the possible contamination of saliva and urine with blood. Patients and methods Twenty three patients with chronic hepatitis B virus infection (positive for HBsAg, hepatitis B e antigen (HBeAg), and HBV-DNA) who attended this hospital for antiviral treatment participated in the study. Two carriers positive for HBsAg and anti-HBe and a healthy subject negative for HBsAg served as controls. Eighteen of the participants provided samples of all the secretions that were requested (group 1). The remaining eight patients (group 2) either failed to provide some of the samples or provided insufficient samples. Patients were issued with receptacles for the specimens and were asked to collect about 3 ml saliva when they woke up in the morning and before brushing their teeth; 50 ml urine voided in the morning; and a sample of semen obtained preferably after abstention from sexual activity for three days. A specimen of serum was obtained on the same day by venepuncture. All specimens were processed within two to three hours after collection. Specimens of saliva and urine were tested for the presence of blood by Hemastix (Ames Division, Miles Laboratories, Slough) before concentration. Samples of semen were allowed to liquefy at room temperature, and the sperm were then separated from the seminal fluid by centrifugation. The saliva, urine, and seminal fluid were next placed in dialysis tubing and concentrated against macrogol 6000 (polyethylene glycol 6000) (BDH Chemicals, Poole). Saliva and seminal fluid were concentrated to 250 IL irrespective of starting volume. Urine was concentrated from a 25 ml starting volume to 250 pl (100-fold concentration). All samples were then stored at -20C. The methods of HBV-DNA extraction, preparation of radiolabelled probes by nick translation, and molecular hybridisation have been described previously.2' 2628 Viral DNA from saliva, urine, and seminal fluid was extracted as for serum. Autoradiographs 1853 1854 BRITISH MEDICAL JOURNAL VOLUME 290 22 JUNE 1985 were obtained by exposure of hybridised DNA on nitrocellulose filters for 24-48 hours. Samples from four patients were also analysed by agarose gel electrophoresis and Southern blotting followed by hybridisation with HBV-DNA labelled with phosphorus-32.26 Results The table shows the incidence of HBV-DNA in saliva, urine, and seminal fluid. HBV-DNA was detected in 55-88% of the samples from the HBeAg positive patients and none from the controls. Moreover, in group 1 at least one sample from every patient was positive for HBV-DNA and all 15 patients were therefore potentially infectious. Nine of these patients provided samples on a second occasion and similar positivity rates were obtained. To confirm that the results obtained were due to HBV-DNA, samples were examined by Southern blotting to investigate the molecular species of the hybridisable DNA. Hybridisation signals in the form of smears covering the size range 2-0-2-8 kb were seen. These corresponded to the heterogeneous population of HBV-DNA associated with virions. The intensity of the autoradiography spots (figure) from all secretions was generally weak and required at least 48 hours of film exposure before becoming visible. This was particularly true for the samples of seminal fluid. Several specimens of saliva and urine, however, produced strong spots within 24 hours. Serum concentrations of HBV-DNA ranged from 8-94xlO-11 to 2 77 x 10-1 mmol/l (186 to 5760 pg/ml). Samples of urine and saliva were examined for blood. None of the samples of urine contained detectable amounts of blood. In the saliva, however, blood was detected in varying amounts (two trace, four small, two moderate, and three large) in 11 of the 17 samples positive for HBV-DNA. Prevalence of HBV-DNA in saliva, urine, and seminal fluid. Figures are proportions of patients (and %) Saliva Urine Seminal fluid Serum Group 1 13/15 (86 6) 9/15 (60) 8/15 (53-3) 15/15 (100) Group 2 2/2 (100) 3/7 (42 8) 5/6 (83) 8/8 (100) Total 15/17 (88/2) 12/22 (545) 13/21 (61-9) 23/23 (100) Controls 0/3 0/3 0/3 0/3 HBsAg HBeAg+ve ,Vcb e oivpa W _,_.f S-m - -v| Composite autoradiograph of nitrocellulose filters bearing samples of saliva, urine, seminal fluid, and serum probed for HBV-DNA using HBV-DNA labeled with phosphorus-32. Black areas indicate presence of HBV-DNA in samples. Intensity of spot indicates amount of HBV-DNA present. Discussion The detection of HBV-DNA in saliva, urine, and seminal fluid by molecular hybridisation indicates the likely presence of hepatitis B virus particles in these secretions and establishes them as potential vehicles of infectivity. Although it has been suspected that non-parenteral transmission of hepatitis B virus may be due to contamination of such secretions with virions, this can now be substantiated by the sensitive technique of molecular hybridisation. Of the concentrated saliva specimens, 87% were positive for HBV-DNA, and in some large amounts were present. The detection of HBV-DNA in some of these may have been due to the presence of oral or gingival lesions as blood was detected in 64% of the samples and this was gross in 31%. In some cases, however, HBV-DNA was detected in saliva with no detectable contamination with blood. Of the concentrated urine specimens, 60% were positive for HBV-DNA and none were contaminated with blood. This prevalence was higher than previous reports of HBsAg in urine,9 10 possibly because of the greater degree of concentration achieved here and the more sensitive technique of molecular hybridisation. Seminal fluid was also found to be positive for HBV-DNA. The degree of concentration varied from twofold to 10-fold depending on the volume of the ejaculate. The presence of HBV-DNA in the above secretions was independent of the amount of HBV-DNA in serum. Although no quantitative measurements were undertaken owing to the difficulty in standardising concentration factors, the amounts detected in the various secretions were small compared with those found in serum. Probably there is transudation or exudation of fluid containing virus from the general circulation into various body fluids rather than active virus replication at the site of secretion. The presence of HBV-DNA in the various secretions was further confirmed by Southern blotting. Positive autoradiographs were obtained from smears of HBV-DNA sequences in the 2-0-2-8 kb region of the gels, where HBV-DNA associated with virions is normally found.26 Most of the men investigated were homosexual, and all of them had at least one sample that was positive for HBV-DNA. It has been postulated that the high rate of transmission of hepatitis B virus in homosexual populations is due to contamination of partners with blood from lacerations in the rectum of the passive partner or cuts on the penis of the active partner.20 Our results prove that this may not be the only mechanism. Contaminated semen or saliva deposited in the rectum during orogenital and oroanal contact might be alternative modes of transmission among homosexual men. Similarly, consorts of heterosexual men are at risk of infection from their partner's secretions. Indeed the potential of saliva and semen to transmit hepatitis B virus infection has been shown experimentally in primates.29 30 References 1 Dienstag JL. Toward the control of hepatitis B. N EnglJ Med 1980;303:874-6. 2 Szmuness W, Much MI, Prince AM, et al. The role of sexual behaviour in the spread of hepatitis B infection. Ann Intern Med 1975;83:489-95. 3 Szmuness W, Stevens CE, Harley EJ, et al. Hepatitis B vaccine demonstration of efficacy in a controlled clinical trial in a high-risk population in the United States. N EnglJ Med 1980;303:833-41. 4 Redeker AG, Mosley JW, Gocke DJ, McKee AP, Pollack W. Hepatitis B immune globulin as a prophylactic measure for spouses exposed to acute type B hepatitis. N Engl_J Med 1975;273:1055-9. 5 Koff RS, Slavin MM, Conneily LJD, Rosen DR. Contagiousness of acute hepatitis B secondary attack rates in household contacts. Gastroenterology 1977;72 :297-300. 6 Inaba N, Ohkawa R, Matsuura A, Kudoh J, Takqumizawa H. Sexual transmission of hepatitis B surface antigen. Infection of husbands by HBsAg carrier state wives. BrJ Vener Dis 1979;55:366-8. 7 Perillo RP, Belb L, Campbell C, et al. Hepatitis B e antigen, DNA polymerase activity, and infection of household contacts with hepatitis B virus. Gastroenterology 1979;76:1319-25. 8 Heathcote J, Cameron CH, Dane DS. Hepatitis B antigen in saliva and semen. Lancet 1974;i:71-5. 9 Sung JL, Chen DS. Hepatitis B surface antigen in saliva, urine and ascites [Abstract]. Hepatogastroenterol 1983;30:183. 10 Villarejos VM, Visona KA, Gutierrez A, Rodriguez A. Role of saliva, urine and feces in the transmission of type B hepatitis. N EnglJ_ Med 1974;291:1375-8. 11 Blainey JD, Earle A, Flewett TH, Williams LKL. Is the urine infective ini serum hepatitis? Lancet 1971;i:797. 12 Tripatzis I, Horst HG. Detection of Australia-SH-antigen in urine. Nature 1971 ;231 :266-7. 13 Linnemann CC, Goldberg S. HBsAg in breast milk. Lancet 1974;ii:155. 14 Boxall EH, Flewett TH, Dane DS, Cameron CH, MacCallum FO, Lee TW. Hepatitis B surface antigen in breast milk. Lancet 1974;ii:1007-8. 15 Darani M, Gerber M. Hepatitis-B antigen in vaginal secretions. Lancet 1974; ii: 1008. 16 Mazzur S. Menstrual blood as a vehicle of Australia antigen transmission. Lancet 1973;i:749. 17 Hoefs JC, Renner IG, Ashcavai M, Redeker AC. Hepatitis B surface antigen in pancreatic and biliary secretions. Gastroenterology 1980;79:191-4. 18 Feinman SV, Berris B, Rebane A, Sinclair JC, Wilson S, Wrobel D. Failure to detect hepatitis B surface antigen in faeces of HBsAg-positive persons. J Infect Dis 1979;140:407-10. 19 Moodie JW, Stannard LM, Kipps A. The problem of the demonstration of hepatitis B antigen in faeces and bile. J ClGn Pathol 1974;27 :693-7. 20 Reiner NE, Judson FN, Bond WW, Francis DP, Petersen NJ. Asymptomatic rectal mucosal lesions and hepatitis B surface antigen at sites of sexual contact in homosexual men with persistent hepatitis B virus infection. Ann Intern Med 1984;96:170-3. 21 Brechot C, Scotto J, Charnay P, et al. Detection of hepatitis B virus DNA in liver and serum. A direct appraisal of the carrier state. Lancet 1981;ii:765-7. 22 Bonino F, Hoyer B, Nelson J, Engle R, Verme G, Gerin J. Hepatitis B
Response from Dr. McGovern
I did not have time to read the entire article since I need to answer many questions tonight, although it looks very interesting...However, I will say that although a virus is recovered in a particular fluid, does not mean that we know it is transmitted in that fluid. This is research that will lead to other behavioral research to better define the importance of these findings.
I would also like to highlight that HCV has been isolated in semen and cervical vaginal fluids. However, sexual transmission of HCV is uncommon among monogamous couples.
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