Detection of Viable but Nonculturable Escherichia coli O157:H7 in Ground Beef by Propidium Monoazide real-time PCR

نویسندگان

  • Jehan Mahmoud
  • Mahmoud Ouf
  • Yuan Yuan
  • Prashant Singh
  • Azlin Mustapha
چکیده

Escherichia coli O157:H7 can enter into a viable but nonculturable (VBNC) state under stress conditions. Pathogens in this dormant state may escape detection if conventional methods are employed, and potentially pose serious threats to human health. Studies have shown that many intervention and preservation processes that are commonly used in the food industry may instead induce a VBNC state rather than kill the intended pathogens. This study aimed to detect whether E. coli O157:H7, an important and dangerous foodborne pathogen, could adapt to the stress caused by lactic acid exposure by entering the VBNC state. A propidium monoazide (PMA) quantitative PCR (qPCR) method was used for detection and quantifi cation of VBNC E. coli O157:H7 cells. The performance of this PMA-qPCR method was assessed using pure culture and ground beef samples inoculated with VBNC E. coli O157:H7 cells. The applied assay could detect as low as 103 CFU/mL VBNC E. coli O157:H7 in pure culture and 4 × 104 CFU/g VBNC cells in ground beef. Results indicate that PMA qPCR could accurately quantify E. coli O157:H7 in a VBNC state. Research Article Detection of Viable but Nonculturable Escherichia coli O157:H7 in Ground Beef by Propidium Monoazide real-time PCR Jehan Mahmoud Mahmoud Ouf1, Yuan Yuan2, Prashant Singh2 and Azlin Mustapha2* 1Food Hygiene Department, Animal Health Research Institute, Dokki, Giza, Egypt 2Food Science Program, University of Missouri, Columbia, Missouri, USA Dates: Received: 06 April, 2017; Accepted: 19 May, 2017; Published: 20 May, 2017 *Corresponding author: Azlin Mustapha, Food Science Program, Division of Food Systems and Bioengineering, 246 William Stringer Wing, Eckles Hall, University of Missouri, Columbia, MO 65211, USA, Tel: +1-573-882-2649; Fax: +1-573-884-7964; E-mail:

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

RNA-Based Detection Does not Accurately Enumerate Living Escherichia coli O157:H7 Cells on Plants

The capacity to distinguish between living and dead cells is an important, but often unrealized, attribute of rapid detection methods for foodborne pathogens. In this study, the numbers of enterohemorrhagic Escherichia coli O157:H7 after inoculation onto Romaine lettuce plants and on plastic (abiotic) surfaces were measured over time by culturing, and quantitative PCR (qPCR), propidium monoazid...

متن کامل

Rapid and Accurate Detection of Bacteriophage Activity against Escherichia coli O157:H7 by Propidium Monoazide Real-Time PCR

Conventional methods to determine the efficacy of bacteriophage (phage) for biocontrol of E. coli require several days, due to the need to culture bacteria. Furthermore, cell surface-attached phage particles may lyse bacterial cells during experiments, leading to an overestimation of phage activity. DNA-based real-time quantitative polymerase chain reaction (qPCR) is a fast, sensitive, and high...

متن کامل

The physiologic state of Escherichia coli O157:H7 does not affect its detection in two commercial real-time PCR-based tests.

Multiplex real-time PCR detection of Escherichia coli O157:H7 is an efficient molecular tool with high sensitivity and specificity for meat safety assurance. The Biocontrol GDS(®) and DuPont Qualicon BAX(®)-RT rapid detection systems are two commercial tests based on real-time PCR amplification with potential applications for quantification of specific E. coli O157:H7 gene targets in enriched m...

متن کامل

Assessments of Total and Viable Escherichia coli O157:H7 on Field and Laboratory Grown Lettuce

Leafy green produce has been associated with numerous outbreaks of foodborne illness caused by strains of Escherichia coli O157:H7. While the amounts of culturable E. coli O157:H7 rapidly decline after introduction onto lettuce in the field, it remains to be determined whether the reduction in cell numbers is due to losses in cell viability, cell injury and a subsequent inability to be detected...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره   شماره 

صفحات  -

تاریخ انتشار 2017