Application of Multiplex PCR Assay for Campylobacter fetus Sub sp. Detection and Differentiation in Bovine Preputial Wash
Abstract: The aim of this
research was to evaluate the detection and differentiation of C. fetus
subspecies through multiplex PCR (mPCR) method from the bovine preputial wash.
The evaluation was done by comparing 2 extraction methods, which was by
commercial kit extraction and boiling method, and also by measuring the effect
of Transport Enrichment Media (TEM) addition as bacterial enrichment. Spiked
bovine preputial washes with 2 extraction methods (commercial kit and boiling),
with and without TEM addition were used as a sample. The results showed that
the highest sensitivity/specificity was by boiling extraction without TEM
addition, which reached 83.3%/100% for all or grouped sample. The results also
supported by Poisson regression analysis which showed the comparison between
sample with and without TEM addition which had 1.3 Incidence Rate Ratio (IRR).
The obtained IRR score indicated that sample without TEM addition had 30%
better result compared to the sample with TEM addition. Furthermore, a
comparison between commercial kit and boiling extraction showed no significant
difference (1.029 IRR). The research concluded that boiling extraction without
TEM addition is the most accurate, fast and economical extraction method
regarding its high sensitivity and specificity value, with less than 24 hours
testing time and without any commercial kit requirements. However, a validation
and verification with field sample were still needed before applied in the
laboratory.
Keywords: Bovine genital
campylobacteriosis; Campylobacter fetus; Cff; Cfv; Multiplex PCR
Penulis: Seruni Agistiana,
Surachmi Setiyaningsih, Agustin Indrawati
Kode Jurnal: jppeternakandd180017
