PCR-ELISA for diagnosis of brucellosis in goat milk.

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作   者：

Gupta VK;  Jyoti Vohra;  Vihan VS;  Ranjeeta Kumari; 

作者机构：

India.;  Uttar Pradesh 281 122;  Central Institute for Research on Goats;  Makhdoom; 

关键词：

diagnostic techniques;  brucellosis;  goats;  diagnosis;  immunodiagnosis;  immunological techniques;  polymerase chain reaction;  Brucella melitensis;  ELISA;  goat milk; 

期刊名称：

The Indian Journal of Animal Sciences

i s s n：

0367-8318

年卷期：

2008 年 78 卷 8 期

页   码：

797-800

页   码：

摘   要：

A simple milk PCR-enzyme-linked immunosorbent assay (PCR-ELISA) was developed to get rid of some of the limitations of conventional microbiological methods in the diagnosis of caprine brucellosis. A 720-bp sequence of a gene that codes for an outer membrane protein (omp31) specific for the Brucella melitensis was amplified. Subsequently, the digoxigenin-labelled amplified product was hybridized with a biotinylated capture probe, which was complementary to the inner part of the amplicon. The hybrid was captured on streptavidin-coated 96 wells plate and detected by using an antidigoxigenin Fab-peroxidase conjugate. The detection limit of the milk PCR-ELISA was 10 fg (two bacterial cells). The sensitivity of milk PCR-ELISA was higher than that of ethidium bromide staining. The PCR-ELISA was 93.75% sensitive and 100% specific, whereas the sensitivity of the milk PCR and serology was 87.5 and 75%, respectively.