The analysis of groEL gene in Salmonella enterica subspecies enterica serovar Typhimurium isolated from avians by PCR-Restriction Fragment Length Polymorphism method

Abstract

Salmonella enterica subspecies enterica serovar Typhimurium causes food-borne outbreaks and systemic diseases in humans and animals. groEL gene (also known as mopA gene in S. Typhimurium), possessing conserved sequence, plays an important role in invasion of bacteria. The purpose of present study was to identify the polymorphism of groEL gene among different avians in different regions by PCR-RFLP method. Fifty two S. Typhimurium isolates (Broiler (n = 13), Layer (n = 12), Duck (n = 5), Goose (n = 5), Sparrow (n = 8), Canary (n = 3), Pigeon (n = 5) and Casco parrot (n = 1). were identified using serotyping as well as multiplex-PCR. Then, amplification of groEL gene performed and amplified products subjected to restriction digestion with BsuRI enzyme. Three RFLP profiles, A, B and C, generated DNA fragments between approximately 100-1,000 bp in size, were observed. The RFLP profile A was observed in 35 (67.3%), profile B in 14 (26.9%) and profile C in 3 (5.77%) of isolates. S. Typhimurium isolates recovered from 13 broilers (two of which profile A, 9 profile B and 2 profile C) and from 8 sparrows (two of which profile A, 5 profile B and 1 profile C) showed all three profiles, but 12 layers and other avians (including Canary (n = 3), Goose (n = 5), Duck (n = 5), Pigeon (n = 5) and Casco parrot (n = 1)) showed profile A. None of these profiles was allotted for a special region. The result of present study showed that S. Typhimurium undergoes genetic mutations in groEL gene under unpleasant milieu in different regions and in different avians. Thus, genetic diversity, despite conserved nature of groEL gene in S. Typhimurium, may exist but it depends on the condition where bacteria have settled. To our knowledge, three RFLP profiles of groEL gene generated by BsuRI restriction enzyme were not reported previously.

PCR-RFLP analysis of fliC, fimH and 16S rRNA genes in Salmonella Typhimurium isolates of varied origin

“…This result shows the potential of ClaI to used in the differentiation of Salmonella Typhimurium isolates. No pattern was allotted for a special region or source which is similar to the observation of Dilmaghani et al (2010) in the fljB gene.…”

“…It has also been shown that genotypic diversity existing in flagellin genes (fliC and fljB encoding phase-1 and phase-2 flagellin, respectively) and FimH protein (fimbrial adhesin) of S. enterica makes them excellent candidates for heterogeneity studies (Guo et al 2007;Jong et al 2010;Kisiela et al 2012). Paradoxically, Dilmaghani et al (2010) found that fljB gene in Salmonella Typhimurium is highly conserved. Based on these findings, we have analyzed the conserved nature or polymorphism of the fliC, 16S rRNA and fimH genes in Salmonella Typhimurium isolates of varied origin using PCR-RFLP.…”

Seleção, caracterização e clonagem dos genes fljB e groEL agonistas dos receptores de reconhecimento de padrão do sistema imune inato das aves

“…Os iniciadores utilizados por Dilmaghani et al (2011) para o gene fljB foram os mesmos adotados no presente estudo. Desta forma, a visualização dos produtos de PCR do gene fljB na eletroforese em gel de agarose com os mesmos tamanhos encontrados por esses autores, confirmaram a presença desse gene nas amostras provenientes de S. Typhimurium.…”