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  IN THIS Article
 ::  Abstract
 ::  Introduction
 ::  Material and methods
 ::  Results
 ::  Discussion
 ::  Acknowledgement
 ::  References
 ::  Article Tables

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Year : 1989  |  Volume : 35  |  Issue : 2  |  Page : 83-5

Comparative study of methods of detection of hepatitis 'B' surface antigen (HBsAg).

Correspondence Address:
V V Parab

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Source of Support: None, Conflict of Interest: None

PMID: 0002621667

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 :: Abstract 

The serum samples were collected from 52 patients of acute viral hepatitis and 235 hospital staff from Kasturba Hospital for Infectious Diseases. HBsAg was detected in their sera by counter-immuno-electrophoresis (CIEP), reverse passive hemogglutination (RPHA) and by micro-enzyme-linked-immunosorbent assay (ELISA) technique. Among the patients, HBsAg was detected in 12 cases (23%) by CIEP, in 18 cases (34%) by RPHA and in 23 patients (45%) by ELISA. In the hospital staff, HBsAg was detected in 4 samples (1.7%) by CIEP, in 8 samples (3.5%) by RPHA and in 32 samples (13.5%) by ELISA. Thus ELISA was found to be the most sensitive technique in detecting HBsAg.

Keywords: Acute Disease, Adolescent, Adult, Comparative Study, Female, Hepatitis B Surface Antigens, analysis,isolation &purification,Hepatitis, Viral, Human, blood,Human, Male, Middle Age,

How to cite this article:
Parab V V, Talawadekar N N, Kale V V, Kamat S A. Comparative study of methods of detection of hepatitis 'B' surface antigen (HBsAg). J Postgrad Med 1989;35:83

How to cite this URL:
Parab V V, Talawadekar N N, Kale V V, Kamat S A. Comparative study of methods of detection of hepatitis 'B' surface antigen (HBsAg). J Postgrad Med [serial online] 1989 [cited 2023 Sep 25];35:83. Available from:

 :: Introduction Top

The detection of Australia antigen has become necessary for diagnosis of Australia antigen positive hepatitis and to find out antigen carrier rate among healthy individuals. A number of methods for detection of HBsAg have been reported, since the demonstration of its relation[2] to viral hepatitis. Most of the methods are directed towards detection of HBsAg. These methods vary in their sensitivity as well as specificity. Some of them are quite simple like, AGD, CIEP, but have very low sensitivity while some of them are more sensitive like RPHA, RIA, ELISA etc. This study was undertaken to compare sensitivity and specificity between CIEP, RPHA, ELISA and to compare the results between patients and hospital staff.

 :: Material and methods Top

The serum samples were collected from 52 patients of suspected acute viral hepatitis and 235 hospital staff from Kasturba Hospital for Infectious Diseases during the period from-January to May 1986 and were preserved at -20C in deep freeze until tested. The sera were then tested for presence of HBsAg by CIEP, RPHA and ELISA. CIEP was done by using agar gel and standard electrophoretic apparatus by method recommended by Kelkar et al.[6] RPHA was done by using Hoechst kits and ELISA was done by using kits for Australia antigen-enzyme II from Abbott Laboratories, Diagnostic division, (Illinois-Chicago).

 :: Results Top

[Table - 1]shows that in 52 patients (37 males and 15 females) of acute viral hepatitis, with their age ranging from 16 years to 55 years, HBsAg was detected in 12 cases (23%) by CIEP, in 18 patients (34%) by RPHA and in 23 patients (45%) by ELISA technique.

[Table - 1]shows that out of 235 hospital staff (130 males and 105 females) their age ranging from 20, years to 60 years, HBsAg was detected in 4 cases (1.7%) by CIEP, in 8 cases (3.5%) by RPHA and in 32 cases (13.5%) by ELISA technique.

 :: Discussion Top

Inspite of a number of techniques developed for detection of HBsAg a few appear to be relatively applicable for large scale screening. Among them CIEP appears to be the method of choice because of its speed and simple procedure.

In the study it way apparent that large number of samples were positive for HBsAg when RPHA was used and still larger number were positive by ELISA. In the present study, the HBsAg carriage rate was 23% by CIEP and 34% by RPHA in patients group. Our results are agreeable with those of Joshi et al.[4] Bapat et al[1] did not find any difference between CIEP and RPHA. Joshi et al.[5] found in their study that RPHA was seven times more sensitive than CIEP. Dutta[3] observed that 20% of patients of viral hepatitis carried HBsAg in their blood by CIEP. In our study it was found that RPHA was one half times more sensitive than CIEP in patients.

In the present study HBsAg was detected in 1.7% of hospital staff by CIEP, 3.5% of staff by RPHA and in 14.5% of staff by ELISA. Thus it can be seen that CIEP is not of much use in detecting low antigenemia found in hospital staff. RPHA was found to be twice more sensitive than CIEP and there was eight fold increase in detection of hospital staff positive for HBsAg when ELISA was used instead of CIEP. Thyagrajan et al[9] detected HBsAg in 4.5 % of hospital employees by CIEP and in 16.5% of employees by RPHA. Ray et al[7] found that 2.6% of blood donors carried HBsAg in their blood by CIEP and HBsAg was present in 11.8% of donors by RPHA. Our results are agreeable with the study of Shanmugam[8] by ELISA technique.

The study clearly shows that ELISA is the most sensitive technique. In most of hospital laboratories, they have switched over to the third generation test like RPHA for screening donors. Now time has come to use still more sensitive test like ELISA to screen donors to prevent transmission of hepatitis to patients and the hospital personnel.

 :: Acknowledgement Top

The authors wish to thank the Medical Superintendent, Kasturba Hospital, Bombay for allowing us to publish this paper.

 :: References Top

1.Bapat, J. P., Kulkarni, S. K., Baxi, A. J. and Kulkarni, K. V.: Hepatitis B antigen detection by reversed passive hemagglutination method-a comparative data. Ind. J. Bled. Res., 65: 766-769, 1977.  Back to cited text no. 1    
2.Blumberg, B. S., Alter, H. J. and Visnich, S.: A "New" antigen in leukema sera. J. Amer. Med. Assoc., 191: 541-546, 1965   Back to cited text no. 2    
3.Dutta, R. N. and Mahammed, G. S.: Incidence of Australia antigen in voluntary and professional blood donors and also in cases of viral hepatitis. Ind. J. Med. Res. 60: 1774-1778, 1972.  Back to cited text no. 3    
4.Joshi, B. N. and Dharmadhikari, C. A.: Hepatitis B antigen in endemic hepatitis in a rural area. Ind. J. Med. Res., 65: 474-477, 1977.  Back to cited text no. 4    
5.Joshi, S. H., Baxi, A. V. and Bhatia, H. M.: Prevalence of hepatitis-B surface antigen by rapid passive haemaglutination inhibition technique. Ind. J. Med. Res., 69: 978-980, 1979.  Back to cited text no. 5    
6.Kelkar, S. S. and Niphadkar, K. B.: Agarose and counterimmunoelectrophoresis. Lancet, 2: 1394-1395, 1974.  Back to cited text no. 6    
7.Ray, S. N., Tawil, G. S., Regis, M. L., Narayanan, R., Rahyalakshmi, K. and Sharma, S. P.: Detection of hepatitis B Surface antigen among blood donors-a comparative study on methods. Ind. J. Med. Res., 65: 478-481, 1977.  Back to cited text no. 7    
8.Shanmugam, J., Balakrishnan, K. G. and Hariprasad, D.: HBs-Ag detection by micro-enzyme linked immunosorbent assay (ELISA) technique. Ind. J. Pathol. Microbiol., 26: 121-125, 1983.  Back to cited text no. 8    
9.Thyagrajan, S. P., Subramanian, S., Sundaravelu, T.. Shivakumar, S., Prasad, P. R. and Thiruvengadam, K. V.: Hepatitis B surface antigen carriers among hospital personnel: (A sero-study in Government General Hospital, Madras). J. Assoc. Phys. Ind., 29: 941-945, 1981.  Back to cited text no. 9    


[Table - 1], [Table - 2]

This article has been cited by
1 Reversed passive hemagglutination test fails to detect HBsAg in a number of HBeAg positive sera
Thammanichanond D, Kunakorn M, Settaudom C, et al.


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2004 - Journal of Postgraduate Medicine
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