Rapid diagnosis of rabies in humans and animals by a dot blot enzyme immunoassay

被引:23
作者
Madhusudana, SN [1 ]
Paul, JPV [1 ]
Abhilash, VK [1 ]
Suja, MS [1 ]
机构
[1] NIMHANS, Dept Neurovirol, Bangalore 560029, Karnataka, India
关键词
rabies; rabies diagnosis; dot blot immunoassay;
D O I
10.1016/j.ijid.2004.02.006
中图分类号
R51 [传染病];
学科分类号
100401 ;
摘要
Objectives: The presently advocated tests for rapid diagnosis of rabies, such as the fluorescent antibody test (FAT) are expensive and require expertise to carry out and interpret the results. In this study, a simple direct dot blot enzyme immunoassay (DIA) has been developed and evaluated to detect the rabies antigen in brain specimens of animals and humans. The utility of this test in the ante-mortem diagnosis of human rabies has also been evaluated. Methods: Brain homogenates of suspected rabid animals (n = 250), humans (n = 16) and clinical samples like saliva (n = 12) and cerebrospinal fluid (CSF) (n = 12) were directly spotted on polyvinylidene difluoride membrane (PVDF) and the absorbed rabies nucleoprotein antigen was detected using biotinylated anti nucleoprotein antibody followed by treatment with streptavidin peroxidase conjugate and color development with diamino benzedine (DAB). Rabies-infected and normal mouse brain homogenates were used as positive and negative controls, respectively. The results of this test were evaluated with fluorescent antibody technique (for brain samples) and mouse inoculation test (for saliva and CSF samples). Results: A distinct dark brown color was seen in the positive control and all positive samples, while there was no color development with either the negative control or the negative samples. The concordance between the fluorescent antibody test (FAT) and dot immunoassay was 98.4% for brain samples, 83.3% for saliva and 91.6% for CSF samples. The specificity of the test was found to be 100%. Conclusions: The dot blot enzyme immunoassay (DIA) test described here is a sensitive, specific and rapid test for the post-mortem diagnosis of rabies in animals and humans. The utility of this test for the ante-mortem diagnosis of rabies needs to be further evaluated. (C) 2004 International Society for Infectious Diseases. Published by Elsevier Ltd. All rights reserved.
引用
收藏
页码:339 / 345
页数:7
相关论文
共 16 条
[1]   Development of Dot-ELISA for the detection of human rotavirus antigen and comparison with RNA-PAGE [J].
Anand, T ;
Raju, TAN ;
Vishnu, C ;
Rao, LV ;
Sharma, G .
LETTERS IN APPLIED MICROBIOLOGY, 2001, 32 (03) :176-180
[2]  
[Anonymous], STRAT CONTR EL RAB A
[3]  
Davis Kennedy L.E, 2000, INFECT DIS NERVOUS S, P401
[4]  
Dean D. J., 1996, P88
[5]  
Dietzschold B., 1996, P175
[6]   Encephalitis caused by a Lyssavirus in fruit bats in Australia [J].
Fraser, GC ;
Hooper, PT ;
Lunt, RA ;
Gould, AR ;
Gleeson, LJ ;
Hyatt, AD ;
Russell, GM ;
Kattenbelt, JA .
EMERGING INFECTIOUS DISEASES, 1996, 2 (04) :327-331
[7]   Dot immunoperoxidase assay for detection of parvovirus B19 antigens in serum samples [J].
Gentilomi, G ;
Musiani, M ;
Zerbini, M ;
Gallinella, G ;
Venturoli, S ;
Manaresi, E .
JOURNAL OF CLINICAL MICROBIOLOGY, 1997, 35 (06) :1575-1578
[8]   USE OF AVIDIN-BIOTIN INTERACTION IN IMMUNOENZYMATIC TECHNIQUES [J].
GUESDON, JL ;
TERNYNCK, T ;
AVRAMEAS, S .
JOURNAL OF HISTOCHEMISTRY & CYTOCHEMISTRY, 1979, 27 (08) :1131-1139
[9]  
ICHHPUJANI RL, 2001, P 4 INT S RAB CONTR, P27
[10]   A DOT ENZYME-LINKED-IMMUNOSORBENT-ASSAY (DOT ELISA) - COMPARISON WITH STANDARD FLUORESCENT-ANTIBODY TEST (FAT) FOR THE DIAGNOSIS OF RABIES IN ANIMALS [J].
JAYAKUMAR, R ;
NACHIMUTHU, K ;
PADMANABAN, VD .
COMPARATIVE IMMUNOLOGY MICROBIOLOGY AND INFECTIOUS DISEASES, 1995, 18 (04) :269-273