Detection of Viable But Non-Culturable State of Escherichia coli O157: H7 Using Reverse Transcription PCR

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KHEZRI MOHAMMAD; REZAEI MASOUD; MOHABBATI MOBAREZ ASHRAF; ZOLFAGHARI MEHDI

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Journal: IRANIAN JOURNAL OF MEDICAL MICROBIOLOGY Year:2019 | Volume:12 | Issue:6 Page(s): 390-398

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Title

Detection of Viable But Non-Culturable State of Escherichia coli O157: H7 Using Reverse Transcription PCR

Author(s)

KHEZRI MOHAMMAD | REZAEI MASOUD | MOHABBATI MOBAREZ ASHRAF | ZOLFAGHARI MEHDI | Issue Writer Certificate

Keywords

Escherichia coli O157:H7Q2

Bacteria viabilityQ1

Reverse Transcriptase PCRQ1

Abstract

Background and Aims: Many bacteria including Escherichia coli may enter into a viable but non-culturable (VBNC) state under unfavorable stresses, which are unable to be detected by culture-based methods. In this study, the use of Reverse Transcription PCR (RT-PCR) for detection of VBNC state of E. coli O157: H7 was investigated. Materials and Methods: Escherichia coli O157: H7 was inoculated in distilled water and 30 percent salt water at room temperature. The cultivability of bacteria was determined using routine culture and colony counting on MacConkey agar. The RT-PCR of 16S rRNA gene involved direct extraction and purification of RNA, DNase I treatment for removing DNA contamination, cDNA synthesis and electrophoresis of PCR products of cDNA was used to detect viable E. coli O157: H7 under studied treatments and was compared with the results of RT-PCR of 16S rRNA gene of heat-killed bacteria. Results: The cultivability of bacteria was maintained during the study period in distilled water; however, the use of 30% NaCl caused the bacteria to be non-cultivable on day 4. The RTPCR of 16S rRNA showed the positive expression of this gene in cultivable and non-cultivable bacteria during the study period, whereas heat-killed bacteria were negative for this gene, which indicated the efficacy of RT-PCR of 16S rRNA in differentiation of alive from dead bacteria. Conclusions: Escherichia coli O157: H7 entered into the VBNC under 30 percent NaCl which can be associated with serious human health problems. RT-PCR can be used to detect bacteria in the VBNC state.

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APA: Copy

KHEZRI, MOHAMMAD, REZAEI, MASOUD, MOHABBATI MOBAREZ, ASHRAF, & ZOLFAGHARI, MEHDI. (2019). Detection of Viable But Non-Culturable State of Escherichia coli O157: H7 Using Reverse Transcription PCR. IRANIAN JOURNAL OF MEDICAL MICROBIOLOGY, 12(6 ), 390-398. SID. https://sid.ir/paper/132537/en

Vancouver: Copy

KHEZRI MOHAMMAD, REZAEI MASOUD, MOHABBATI MOBAREZ ASHRAF, ZOLFAGHARI MEHDI. Detection of Viable But Non-Culturable State of Escherichia coli O157: H7 Using Reverse Transcription PCR. IRANIAN JOURNAL OF MEDICAL MICROBIOLOGY[Internet]. 2019;12(6 ):390-398. Available from: https://sid.ir/paper/132537/en

IEEE: Copy

MOHAMMAD KHEZRI, MASOUD REZAEI, ASHRAF MOHABBATI MOBAREZ, and MEHDI ZOLFAGHARI, “Detection of Viable But Non-Culturable State of Escherichia coli O157: H7 Using Reverse Transcription PCR,” IRANIAN JOURNAL OF MEDICAL MICROBIOLOGY, vol. 12, no. 6 , pp. 390–398, 2019, [Online]. Available: https://sid.ir/paper/132537/en

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