Detection of multiple waterborne pathogens using microsequencing arrays

J Appl Microbiol. 2013 Feb;114(2):564-73. doi: 10.1111/jam.12073. Epub 2012 Dec 27.

Abstract

Aims: A microarray was developed to simultaneously detect Cryptosporidium parvum, Cryptosporidium hominis, Enterococcus faecium, Bacillus anthracis and Francisella tularensis in water.

Methods and results: A DNA microarray was designed to contain probes that specifically detected C. parvum, C. hominis, Ent. faecium, B. anthracis and F. tularensis. The microarray was then evaluated with samples containing target and nontarget DNA from near-neighbour micro-organisms, and tap water spiked with multiple organisms. Results demonstrated that the microarray consistently detected Ent. faecium, B. anthracis, F. tularensis and C. parvum when present in samples. Cryptosporidium hominis was only consistently detected through the use of shared probes between C. hominis and C. parvum.

Conclusions: This study successfully developed and tested a microarray-based assay that can specifically detect faecal indicator bacteria and human pathogens in tap water.

Significance and impact of the study: The use of indicator organisms has become a practical solution for monitoring for water quality. However, they do not always correlate well with the presence of many microbial pathogens, thus necessitating direct monitoring for most pathogens. This microarray can be used to simultaneously detect multiple organisms in a single sample. More importantly, it can provide occurrence information that may be used in assessing potential exposure risks to waterborne pathogens.

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Bacillus anthracis / genetics
  • Bacillus anthracis / isolation & purification
  • Bacteria / isolation & purification*
  • Cryptosporidium / genetics
  • Cryptosporidium / isolation & purification*
  • Cryptosporidium parvum / genetics
  • Cryptosporidium parvum / isolation & purification
  • Enterococcus faecium / genetics
  • Enterococcus faecium / isolation & purification
  • Feces
  • Francisella tularensis / genetics
  • Francisella tularensis / isolation & purification
  • Oligonucleotide Array Sequence Analysis / methods*
  • Oligonucleotide Probes
  • Sequence Analysis, DNA
  • Water / parasitology*
  • Water Microbiology*

Substances

  • Oligonucleotide Probes
  • Water