Evaluation of indigenous milk ELISA with m-culture and m-PCR for the diagnosis of Bovine Johne’s disease (BJD) in lactating Indian dairy cattle

Present study is the first attempt to evaluate an indigenous milk ELISA with milk culture, standardize milk PCR, estimate lacto-prevalence of Map and genotype Map DNA from milk samples in few Indian dairy herds. In all 115 cows were sampled from 669 lactating cows in six dairy herds from three distr...

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Bibliographic Details
Published in:Research in veterinary science 2008-02, Vol.84 (1), p.30-37
Main Authors: Sharma, G., Singh, S.V., Sevilla, I., Singh, A.V., Whittington, R.J., Juste, R.A., Kumar, S., Gupta, V.K., Singh, P.K., Sohal, J.S., Vihan, V.S.
Format: Article
Language:English
Subjects:
Animals
Bacteriological Techniques - veterinary
Buffalo
Cattle
cattle diseases
Cattle Diseases - diagnosis
Cattle Diseases - microbiology
Dairy cattle
dairy cows
dairy herds
Dairying
Deoxyribonucleic acid
Disease
disease detection
disease prevalence
DNA
enzyme-linked immunosorbent assay
Enzyme-Linked Immunosorbent Assay - veterinary
Female
Genetic testing
genotype
Goats
in vitro culture
India
IS 1311 PCR–REA
Johne’s disease
Lactating cows
Lactation
Microbiology
Milk
Milk - chemistry
Milk - microbiology
Milk culture
Milk ELISA test
Milk PCR
molecular epidemiology
Mycobacterium avium subsp. paratuberculosis
Mycobacterium avium subsp. paratuberculosis - isolation & purification
paratuberculosis
Paratuberculosis - diagnosis
pathogen identification
polymerase chain reaction
Polymerase Chain Reaction - veterinary
reference standards
screening
seroprevalence
Veterinary medicine
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Summary:Present study is the first attempt to evaluate an indigenous milk ELISA with milk culture, standardize milk PCR, estimate lacto-prevalence of Map and genotype Map DNA from milk samples in few Indian dairy herds. In all 115 cows were sampled from 669 lactating cows in six dairy herds from three districts of North India. Fifty milk samples (four herds) were screened by three tests (milk culture, m-ELISA and m-PCR). Lacto-prevalence of Map in four dairy herds was 84.0% (50.0% in fat and 62.0% in sediment). Screening of both fat and sediment increased the sensitivity of culture. Colonies appeared between 45 and 120 DPI. In indigenous m-ELISA, protoplasmic antigen derived from native Map ‘Bison type’ strain of goat origin was used. Screening of 115 lactating cows by m-ELISA (‘herd screening test’) detected 32.1% positive lactating cows (lacto-prevalence). Sensitivity of ELISA was 28.5% and 42.8% in single point cutoff and S/P ratio, respectively. Lacto-prevalence of JD was high in dairy herds (66.6–100.0% by culture and 20.0–50.0% by m-ELISA). DDD farm, Mathura had very high (95.8%) and moderate prevalence of Map and lacto-antibodies, respectively. All cows were clinically suffering from JD. Specific IS 900 PCR was standardized in decontaminated fat and sediment of milk samples. DNA isolated from decontaminated pellets was amplified and characteristic 229 bp band was confirmatory for Map. Of the 50 milk samples, 6.0% were positive in m-PCR. The test needs further standardization. Map DNA were genotyped as Map ‘Bison type’ by IS 1311 PCR–REA. Of the three tests, milk culture was most sensitive followed by m-ELISA and m-PCR. Map DNA isolated from milk samples of dairy cattle were first time genotyped as Map, ‘Bison type’ in India. High prevalence of Map in milk of dairy herds, posed major health hazard for calves and human beings.
ISSN:0034-5288
1532-2661
DOI:10.1016/j.rvsc.2007.03.014