Real-time PCR detection of male-specific coliphages.
Male-specific coliphages are often used as indicators of contamination by enteric viruses. These phages can be detected in water samples by plaque assays and by polymerase chain reaction. In this study, the M13 coliphage was used to develop a real-time PCR assay for the detection of male-specific DNA coliphages. The real-time PCR was found to have a reaction efficiency of 1.45 and detection limit of 10(-3) plaque forming units per reaction mix. Repeated amplification and melting curve analyses demonstrated high specificity and reproducibility of the real-time assay. Quantitative detection with the real-time PCR should allow rapid assessment of the level of viral contamination in water.
Yong
S F Y
SF
School of Arts and Sciences, Monash University Malaysia, Jalan Lagoon Selatan, Bandar Sunway, Petaling Jaya 46150, Selangor, Malaysia. stacey.yong@artsci.monash.edu.my
Ngeow
Y F
YF
Tong
Y K
YK
Ong
J T
JT
D017104:Bacteriophage M13 / Q000235:genetics* / Q000302:isolation & purification*; D004279:DNA, Viral / Q000032:analysis*; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D014871:Water Microbiology
D004279:DNA, Viral