ABSTRACT
Introduction@#PCR to detect and amplificate the virulence genes of STEC is specific and more sensitive, however, it takes five to six hours for whole test procedure and requires special lab instruments such as thermocycler. Loop-mediated isothermal amplification is a simple, rapid, specific and cost-effective nucleic acid amplification method by using four to six primers when compared to PCR, nucleic acid sequence-based amplification, self-sustained sequence replication and strand displacement amplification.@*Goal@#Detection and comparison of STEC by PCR and LAMP@*Materials and Methods@#In our study, we analyzed comparison of PCR and LAMP results on standard strain used quality control strain solution which diluted 1pg/µL DNA, 10 pg/µL DNA, 100 pg/µL DNA, 1 ng/μL DNA, 10 ng/μL DNA, and 100 ng/μL DNA concentration from LB agar cultures. @*Research ethics@#Permission to submit the survey was granted by the Ethics Review Committee of the MNUMS and the survey was conducted in accordance with the rules and regulations.@*Result@#Sensitivity of Stx1 and stx2 genes in PCR results are positive in 10 pg/µL DNA solution and negative in 1pg/µL DNA. In LAMP test results showed that positive for all concentration. It shows that LAMP method sensitivity is 10 times more than PCR. @*Conclusion@#It shows that LAMP method sensitivity is 10 times more than PCR. All in allLAMP test is cost effective test with sensitive for detection STEC.
ABSTRACT
Introduction@#PCR to detect and amplificate the virulence genes of STEC is specific and more sensitive, however, it takes five to six hours for whole test procedure and requires special lab instruments such as thermocycler. Loop-mediated isothermal amplification is a simple, rapid, specific and cost-effective nucleic acid amplification method by using four to six primers when compared to PCR, nucleic acid sequence-based amplification, self-sustained sequence replication and strand displacement amplification.@*Materials and Methods@#In our study, we analyzed comparison of PCR and LAMP results on standard strain used quality control strain solution which diluted 1pg/µL DNA, 10 pg/µL DNA, 100 pg/µL DNA, 1 ng/μL DNA, 10 ng/μL DNA, and 100 ng/μL DNA concentration from LB agar cultures. @*Research ethics@#Permission to submit the survey was granted by the Ethics Review Committee of the MNUMS and the survey was conducted in accordance with the rules and regulations. @*Goal@#Detection and comparison of STEC by PCR and LAMP@*Result@#Sensitivity of Stx1 and stx2 genes in PCR results are positive in 10 pg/µL DNA solution and negative in 1pg/µL DNA. In LAMP test results showed that positive for all concentration. It shows that LAMP method sensitivity is 10 times more than PCR. @*Conclusion@#It shows that LAMP method sensitivity is 10 times more than PCR. All in allLAMP test is cost effective test with sensitive for detection STEC.
ABSTRACT
Introduction@#Foodborne diseases are a major public health concern worldwide. The report, which estimates the burden of foodborne diseases – states that each year as many as 600 million, or almost 1 in 10 people in the world, fall ill after consuming contaminated food. Of these, 420 000 people die, including 125 000 children under the age of 5 years. The 20.3% of diarrhea and 27.5% of die caused by contaminated foods are diarrheagenic Escherichia coli (DEC).@*Aim@#To identify of DEC and determine their antibiotic resistance from ready-to-eat salads@*Material and Methods@#A total of 40 bagged salad mix samples were collected from food markets in Ulaanbaatar, Mongolia. Escherichia coli (E.coli) strains were determined on the basis of MNS 6308:2012 standard and confirmed by polymerase chain reaction (PCR) in samples. DEC was identified using multiplex PCR. Bacterial susceptibility to antimicrobial agents determined by the Kirby Bauer disk diffusion method.@*Results@#Our results showed the presence of E. coli in 19 samples (47.5%). DEC isolates identified by multiplex PCR were defined as follows: the presence of eae and bfp for EPEC, the presence of lt for ETEC, the presence of ipaH for EIEC, the presence of stx1 and stx2 for EHEC, the presence of aap and aggR for EAEC, and the presence of daaE for DAEC. The multiplex PCR assays detected EHEC 6 (31.6%), EPEC 5 (26.3%), EIEC 1 (5.3%). EAEC and ETEC were not detected in samples. The E.coli isolates were 73.7% resistant to chloramphenicol as the first choice of treatment of diarrhea and high resistance (68.4-94.7%) to the cephalosporins. In our country, cephalosporins are widely used in medical practice for the treatment of infectious diseases.@*Conclusion@#In this study, about half of ready-to-eat salads are contaminated with E. coli. The three types (EHEC, EPEC, EIEC) of DEC pathotypes were identified in the ready-to-eat salads and high prevalent of antimicrobial resistance. Future research is required to track the contamination sources and develop appropriate steps that should be taken by industry and retailers to reduce microbial contamination in ready-to-eat salads.