Comparison of PCR-RFLP, ribotyping and ERIC-PCR for typing<i>Bacillus anthracis</i>and<i>Bacillus cereus</i>strains

Strains of species in the Bacillus cereus group are potentially enterotoxic. Thus, the detection of all B. cereus group strains is important. As 16S ribosomal DNA sequence analysis cannot adequately differentiate species of the B. cereus group, we explored the potential of the groEL gene as a phylogenetic marker. A phylogenetic analysis of the groEL sequences of 78 B. cereus group strains revealed that the B. cereus group strains were split into two major clusters, one including six B. mycoides and one B. pseudomycoides (cluster II) and the other including two B. mycoides and the rest of the B. cereus group strains (cluster I). Cluster I was further differentiated into two subclusters, Ia and Ib. The sodA gene sequences of representative strains from different clusters were also compared. The phylogenetic tree constructed from the sodA sequences showed substantial similarity to the tree constructed from the groEL sequences. Based on the groEL sequences, a PCR assay for detection and identification of B. cereus group strains was developed. Subsequent restriction fragment length polymorphism (RFLP) analysis verified the PCR amplicons and the differentiation of the B. cereus group strains. RFLP with MboI was identical for all the B. cereus group strains analyzed, while RFLP with MfeI or PstI classified all B. cereus and B. thuringiensis strains into two groups. All cluster II B. mycoides and B. pseudomycoides strains could be discriminated from other B. cereus group bacteria by restriction analysis with TspRI.

show abstract

“…vrrA PCR products were amplified with primers GPR6 and GPR7 (Table 3). vrrA RFLP was done by the method of Shangkuan et al (45).…”

Section: Bacterialmentioning

confidence: 99%

PCR Assay of the groEL Gene for Detection and Differentiation of Bacillus cereus Group Cells

Chang

Shangkuan

Lin

et al. 2003

Appl Environ Microbiol

View full text Add to dashboard Cite

show abstract

“…Therefore, it was possible to consider the analysis of polymorphism by ERIC-PCR, a good tool capable of discriminating these two distinct groups. The analysis and genetic diversity by methods based on repetitions (REP-PCR, BOX-PCR, and ERIC-PCR) have been considered a good discriminatory tool among the isolates of Bacillus (Shangkuan et al, 2000;da Silva and Valicence, 2013;Kumar et al, …”

Section: Resultsmentioning

confidence: 99%

Research Article Genetic diversity of Bacillus sp producers of amylase isolated from the soil.

Xavier¹,

Lima

Oliveira

et al. 2017

Genet. Mol. Res.

View full text Add to dashboard Cite

“…There are several techniques based on analysis of 16S rRNA gene including restriction fragment length polymorphism (RFLP) and sequencing. The RFLP analysis of amplified DNA has been proposed as valuable tool for epidemiological typing of microorganisms (Shangkuan et al 2000;Gray et al 2001).…”

Section: Introductionmentioning

confidence: 99%

Antibiotic resistance and genetic diversity in water-borne Enterobacteriaceae isolates from recreational and drinking water sources

Kumar

Tripathi

Garg

2012

Int. J. Environ. Sci. Technol.

View full text Add to dashboard Cite

A total of 240 water-borne bacteria including 72 Escherichia coli, 83 Enterobacter, 30 Klebsiella, 36 Salmonella and 19 Shigella spp. isolates from drinking and recreational water sources were assessed for antibiotic resistance and genetic diversity. Escherichia coli (88.89 %) and species of Enterobacter (86.75 %), Klebsiella (83.33 %) and Salmonella (100 %) were resistant to cefadroxil, while [94 % Shigella spp. were resistant to cefaclor and cefuroxime. Ofloxacin was the most effective antibiotic against isolates of all the genera. Multiple antibiotic resistance index identified dug well, pond and piped water supplies as high risk sources of enteric pathogens. Random amplified polymorphic DNA analysis and restriction fragment length polymorphism of amplified 16S rRNA gene were studied for genetic relatedness of Enterobacteriaceae isolates. Primer P1254 identified 10, 16, 4, 4 and 1 distinct random amplified polymorphic DNA group(s) of E. coli, Enterobacter, Klebsiella, Salmonella and Shigella species, respectively. Unlike random amplified polymorphic DNA, restriction fragment length polymorphism using AluI and HaeIII could not segregate isolates in different genetic profiles. 16S rRNA gene of three Enterobacter spp. strains from different sources with similar restriction fragment length polymorphism but different random amplified polymorphic DNA patterns was sequenced, and identified as Enterobacter hormaechei strains skg0061, 0062 and 0063. The sequence information has been submitted to GenBank (HQ322393-95). Biochemically similar but genetically diverse Enterobacteriaceae members from drinking and recreational water sources exhibited varying antibiotic sensitivity. Contamination of water sources with such multiple antibiotic-resistant enteric pathogens poses threat to human health.

show abstract

Comparison of PCR-RFLP, ribotyping and ERIC-PCR for typingBacillus anthracisandBacillus cereusstrains

Cited by 54 publications

References 40 publications

PCR Assay of the groEL Gene for Detection and Differentiation of Bacillus cereus Group Cells

PCR Assay of the groEL Gene for Detection and Differentiation of Bacillus cereus Group Cells

Research Article Genetic diversity of Bacillus sp producers of amylase isolated from the soil.

Antibiotic resistance and genetic diversity in water-borne Enterobacteriaceae isolates from recreational and drinking water sources

Contact Info

Product

Resources

About