Comparative study on cultural and molecular methods for detection

Comparative study on cultural and molecular methods for detection of Brucella melitensis in sheep and goat

Title: Comparative study on cultural and molecular methods for detection of Brucella melitensis in sheep and goat

Authors: KB Patel, SI Patel, BK Patel, HC Chauhan, AG Bhagat and BS Chandel

Source: Ruminant Science (2017)-6(2):309-314.

Cite this reference as: Patel KB, Patel SI, Patel BK, Chauhan HC, Bhagat AG and Chandel BS (2017). Comparative study on cultural and molecular methods for detection of Brucella melitensis in sheep and goat. Ruminant Science 6(2):309-314.

Abstract

Brucellosis due to Brucella melitensis is widespread in India and is considered to be the major cause of abortion in small ruminants incurring severe economic loss. In the present study, a total 170 clinical samples of vaginal swab, placental cotyledons, aborted foetal organs and stomach contents, hygroma and orchitis fluid, whole blood and milk were collected but unable to cultural isolation of the Brucella organism. Among the three different genus specific primer pairs used for amplification by PCR, Only B4/B5 and JPF/JPR primer pair detected three samples positive for Brucella DNA from 170 clinical samples.  Proper selection of primer pair is essential, as it was affected by the presence of genomic DNA and finally PCR efficiency. Out of combination of three species specific primers pairs (omp31, ORF A0503 and B. abortus+ IS711) used, only omp31 primer pair produced desired amplicon of 720bp in three clinical samples. This indicated that these animals were infected with Brucella melitensis. Comparatively PCR was found more suitable method for detection Brucella in clinical samples as compared to cultural methods because 3 samples were found positive by PCR which were negative in cultural isolation.

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