TY  - JOUR
T1  - PCR-Based Detection of <I>Aeromonas</I> from Milk Samples
AU - , Porteen, K. AU - , R.K. Agarwal AU - , K.N. Bhilegaonkar 
JO  - Journal of Food Technology
VL  - 4
IS  - 2
SP  - 111
EP  - 115
PY  - 2006
DA  - 2001/08/19
SN  - 1684-8462
DO  - jftech.2006.111.115
UR  - https://makhillpublications.co/view-article.php?doi=jftech.2006.111.115
KW  - A. hydriphila
KW  -milk
KW  -sensitivity
KW  -specificity
AB  - A set of primers targeting 16S rRNA gene and aerolysin gene was employed to standardize PCR assay for detection of <I>Aeromonas</I> from milk samples. The primers used were found to be highly specific for <I>Aeromonas</I> spp and did not give positive result with other Gram positive and Gram-negative bacteria. The minimum detection level of PCR was found to be 10<SUP>2</SUP> cells <sup>-1</SUP> and 10<SUP>4</SUP>cells <sup>-1</SUP> in case of 16S rRNA and aerolysin gene targeted assay, respectively. Suitability of the enrichment broth (Alkaline peptone water-cephalothin,  APW-C)  when  tested  to  detect  <I>Aeromonas</I>  from the spiked samples gave good results on direct  usage of the broth for template preparation without any subsequent treatment. The kinetics of the spiking study indicated that a minimum of 24 hrs enrichment was required for the detection of <I>Aeromonas</I> by cultural and PCR method. Among two PCR assays detection limits achieved by PCR targeting16S rRNA gene were better  than aerolysin gene PCR assay. The results were comparable to cultural method. Examination of 50  milk samples(Pooled samples) revealed two samples to be positive by cultural method and PCR targeting 16S r RNA.Thus the standardized single –step enrichment PCR protocol holds promise as a reliable and rapid method for the detection of <I>Aeromonas</I> from milk samples.
ER  -