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Purification and Visualization of Lipopolysaccharide from Gram-negative Bacteria by Hot Aqueous-phenol Extraction
Published on: May 28, 2012
Molecular insight of putative pathogenicity markers with ESBL genes and lipopolysaccharide in Laribacter
Krishna M Raja1, Asit Ranjan Ghosh
1Centre for Infectious Diseases and Control, School of Bio Sciences and Technology, VIT University, Vellore, 632014, Tamil Nadu, India.
Abstract:
Laribacter hongkongensis is an emerging bacterial pathogen causing gastroenteritis and traveller's diarrhoea. However, pathogenicity of L. hongkongensis has not yet been properly understood. We therefore, investigated putative pathogenicity markers like elt, est, stx1, stx2, eae, eaf, EAgg, bfpA, Int I, Int II, Int III and cnf respectively in strains received from Queen Mary Hospital, Hong Kong University. Above genes were commonly found among pathogenic member genera of Enterobacteriaceae-causing diarrhoea. Received strains were confirmed microbiologically and by 16S ribosomal RNA (rRNA) sequencing. Cultivability was examined using 23 different commercially available microbial growth media followed by antibiotic susceptibility test, lipopolysaccharide (LPS) extraction and raising antisera in rabbit against heat-killed L. hongkongensis. Moreover, Escherichia coli classification genes chuA, yjaA, TspE4 and extended spectrum beta lactamase (ESBL) genes like TEM, SHV, OXA, CTXM, CTXM1 and CTXM9 were also examined by PCR assay. Results showed the possession of eae, bfpA, Int and CTXM9 respectively for putative virulence. Furthermore, purity of extracted LPS was confirmed by HPLC, and raised serum was found useful in diagnosis of LPS; bacterium thus can be employed for immunodiagnostics.
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