Detection of viable Mycobacterium avium subspecies paratuberculosis in powdered infant formula by phage-PCR and confirmed by culture

Int J Food Microbiol. 2016 Jan 4:216:91-4. doi: 10.1016/j.ijfoodmicro.2015.09.011. Epub 2015 Sep 21.

Abstract

Surveys from different parts of the world have reported that viable Mycobacterium avium subsp. paratuberculosis (MAP) can be cultured from approximately 2% of samples of retail pasteurised milk samples. Pasteurised milk is used for the production of powdered infant formula (PIF) and therefore there is a concern that MAP may also be present in these products. Several studies have previously reported the detection of MAP in PIF using PCR-based assays. However, culture-based surveys of PIF have not detected viable MAP. Here we describe a phage amplification assay coupled with PCR (page-PCR) that can rapidly detect viable MAP in PIF. The results of a small survey showed that the phage-PCR assay detected viable MAP in 13% (4/32) of PIF samples. Culture detected viable MAP in 9% (3/32) PIF samples, all of which were also phage-PCR positive. Direct IS900 PCR detected MAP DNA in 22% (7/32) of PIF samples. The presence of viable MAP in PIF indicates that MAP either survived PIF manufacturing or that post-production contamination occurred. Irrespective of the route of MAP contamination, the presence of viable MAP in PIF is a potential public health concern.

Keywords: PCR; Paratuberculosis; Pasteurised milk; Phage; Powdered infant formula; Rapid methods.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacteriophages / genetics
  • DNA, Bacterial / genetics*
  • Humans
  • Infant
  • Infant Formula / microbiology*
  • Infant, Newborn
  • Milk / microbiology*
  • Mycobacterium avium subsp. paratuberculosis / genetics
  • Mycobacterium avium subsp. paratuberculosis / isolation & purification*
  • Polymerase Chain Reaction / methods

Substances

  • DNA, Bacterial