Klebsiella Pneumoniae Research Paper

OCCURRENCE OF SHV GENE AND ANTIBIOGRAM OF KLEBSIELLA PNEUMONIAE INFECTION IN SWISS ALBINO MICE COLONY

M. Thangapandiyan*, S.P. Preetha, S. SureshKannan and T. Mohanapriya
Department of Veterinary Pathology, Madras Veterinary College, Tamil Nadu Veterinary and Animal Sciences University, Chennai – 600 051, India.
*Corresponding author Email: [e-mail]
ABSTRACT:
An epizootic causing high morbidity and high mortality in Swiss albino inbred mice over a period of two weeks caused by Klebsiella pneumoniae is reported. Diagnosis was based on clinical signs, histopathological, bacteriological evaluation of the lung, kidney and intestinal tissue and multiplex PCR. Biochemical tests were positive for citrate, nitrate, urease, lysine decarboxylase,

Equal volume of chloroform:isoamyl alcohol (24:1 v/v) was added, centrifuged at 8000 g for 15 min. The upper layer was used as crude DNA. Amplification reactions for SHV gene was performed in a 25 µl of 10X PCR reaction buffer with Mgcl2 (15mM), 0.5 µl (200µM), deoxynucleoside triphosphate mix (dNTPs, 10mM), 0.5 µl of the primer (10pm/µl) with 0.5 µl (3U/ µl) Taq DNA polymerase (Bangalore Genie, India). Five microlitres of the template DNA preparation was added to the reaction mixture and was carried out on My Cycler DNA thermal cycler (BioRad, Australia). After initial denaturation at 94˚C for 2 min, reaction was performed in a 3 step cycling condition of denaturation for 25 cycles of 1 min at 95˚C, annealing at 52˚C for 55 sec and extension at 72˚C for 45 sec. Final extension was carried out at 72˚C for 5 min. The resulting products were analysed by electrophoresis with 1.5% agarose gels in Tris-borate-EDTA buffer (TBE; Gibco, NY, USA). The gels were stained with ethidium bromide and the band observed at the desired position was photographed on an ultraviolet light transilluminator