title: A simple and novel modification of comet assay for determination of bacteriophage mediated bacterial cell lysis.
abstract: The comet assay is the widely used method for in vitro toxicity testing which is also an alternative to the use of animal models for in vivo testing. Since, its inception in 1984 by Ostling and Johansson, it is being modified frequently for a wide range of application. In spite of its wide applicability, unfortunately there is no report of its application in bacteriophages research. In this study, a novel application of comet assay for the detection of bacteriophage mediated bacterial cell lysis was described. The conventional methods in bacteriophage research for studying bacterial lysis by bacteriophages are plaque assay method. It is time consuming, laborious and costly. The lytic activity of bacteriophage devours the bacterial cell which results in the release of bacterial genomic material that gets detected by ethidium bromide staining method by the comet assay protocol. The objective of this study was to compare efficacy of comet assay with different assay used to study phage mediated bacterial lysis. The assay was performed on culture isolates (N=80 studies), modified comet assay appear to have relatively higher sensitivity and specificity than other assay. The results of the study showed that the application of comet assay can be an economical, time saving and less laborious alternative to conventional plaque assay for the detection of bacteriophage mediated bacterial cell lysis.
authors.affiliation: Environmental Virology Cell (EVC), Council of Scientific and Industrial Research-National Environmental Engineering Research Institute (CSIR-NEERI), Nehru Marg, Nagpur 440020, Maharashtra, India. Electronic address: kskhairnar@gmail.com.
authors.forename: Krishna
authors.initials: K
authors.lastname: Khairnar
authors.affiliation: Environmental Virology Cell (EVC), Council of Scientific and Industrial Research-National Environmental Engineering Research Institute (CSIR-NEERI), Nehru Marg, Nagpur 440020, Maharashtra, India. Electronic address: swamukh1985in@rediffmail.com.
authors.forename: Swapnil
authors.initials: S
authors.lastname: Sanmukh
authors.affiliation: Environmental Virology Cell (EVC), Council of Scientific and Industrial Research-National Environmental Engineering Research Institute (CSIR-NEERI), Nehru Marg, Nagpur 440020, Maharashtra, India. Electronic address: rajshri1022@gmail.com.
authors.forename: Rajshree
authors.initials: R
authors.lastname: Chandekar
authors.affiliation: Environmental Virology Cell (EVC), Council of Scientific and Industrial Research-National Environmental Engineering Research Institute (CSIR-NEERI), Nehru Marg, Nagpur 440020, Maharashtra, India. Electronic address: wn_paunikar@neeri.res.in.
authors.forename: Waman
authors.initials: W
authors.lastname: Paunikar
mesh_terms: D001433:Bacteriolysis; D001435:Bacteriophages / Q000254:growth & development*; D020552:Comet Assay / Q000379:methods*; D004269:DNA, Bacterial / Q000032:analysis*; D012680:Sensitivity and Specificity
chemical_list: D004269:DNA, Bacterial
keywords: Bacteriophage; Comet assay; In vitro toxicity; Plaque assay; Virology research