Detection of multiple waterborne pathogens using microsequencing arrays

• Published in: Journal of applied microbiology Vol. 114; no. 2; pp. 564 - 573
• Main Authors: Brinkman, N.E., Francisco, R., Nichols, T.L., Robinson, D., Schaefer, F.W., Schaudies, R.P., Villegas, E.N.
• Format: Journal Article
• Language: English
• Published: Oxford Blackwell 01-02-2013; Oxford University Press
• Subjects: Bacillus anthracis - genetics; Bacillus anthracis - isolation & purification; Bacteria; Bacteria - isolation & purification; Biological and medical sciences; Cryptosporidium - genetics; Cryptosporidium - isolation & purification; Cryptosporidium parvum - genetics; Cryptosporidium parvum - isolation & purification; Drinking water; Enterococcus faecium - genetics; Enterococcus faecium - isolation & purification; environmental/recreational water; Feces; Food industries; Food microbiology; Francisella tularensis - genetics; Francisella tularensis - isolation & purification; Fundamental and applied biological sciences. Psychology; microarray; microbial contamination; Microbiology; Oligonucleotide Array Sequence Analysis - methods; Oligonucleotide Probes; Pathology; Sequence Analysis, DNA; Water - parasitology; Water Microbiology; Water pollution; water quality; Drinking water; Analysis method; environmental/recreational water; Applied microbiology; Pathogenic; Water quality; microbial contamination; Control method; Detection; Microarray; Biological contamination
• ISSN: 1364-5072; 1365-2672
• DOI: 10.1111/jam.12073

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.