Evaluation of simplified DNA extraction methods for emm typing of group A streptococci.

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dc.contributor.authorJose, J J Men_US
dc.contributor.authorBrahmadathan, K Nen_US
dc.date.accessioned2006-04-12en_US
dc.date.accessioned2009-05-28T09:06:40Z
dc.date.available2006-04-12en_US
dc.date.available2009-05-28T09:06:40Z
dc.date.issued2006-04-12en_US
dc.description.abstractSimplified methods of DNA extraction for amplification and sequencing for emm typing of group A streptococci (GAS) can save valuable time and cost in resource crunch situations. To evaluate this, we compared two methods of DNA extraction directly from colonies with the standard CDC cell lysate method for emm typing of 50 GAS strains isolated from children with pharyngitis and impetigo. For this, GAS colonies were transferred into two sets of PCR tubes. One set was preheated at 94 degrees C for two minutes in the thermal cycler and cooled while the other set was frozen overnight at -20 degrees C and then thawed before adding the PCR mix. For the cell lysate method, cells were treated with mutanolysin and hyaluronidase before heating at 100 degrees C for 10 minutes and cooling immediately as recommended in the CDC method. All 50 strains could be typed by sequencing the hyper variable region of the emm gene after amplification. The quality of sequences and the emm types identified were also identical. Our study shows that the two simplified DNA extraction methods directly from colonies can conveniently be used for typing a large number of GAS strains easily in relatively short time.en_US
dc.description.affiliationDepartment of Clinical Microbiology, Christian Medical College, Vellore - 632 004, Tamil Nadu, India.en_US
dc.identifier.citationJose JJ, Brahmadathan KN. Evaluation of simplified DNA extraction methods for emm typing of group A streptococci. Indian Journal of Medical Microbiology. 2006 Apr; 24(2): 127-30en_US
dc.identifier.urihttps://imsear.searo.who.int/handle/123456789/53445
dc.language.isoengen_US
dc.source.urihttps://www.ijmm.orgen_US
dc.subject.meshAntigens, Bacterial --geneticsen_US
dc.subject.meshBacterial Outer Membrane Proteins --geneticsen_US
dc.subject.meshBacterial Proteins --geneticsen_US
dc.subject.meshBacterial Typing Techniques --methodsen_US
dc.subject.meshChilden_US
dc.subject.meshChild, Preschoolen_US
dc.subject.meshDNA, Bacterial --analysisen_US
dc.subject.meshFreezingen_US
dc.subject.meshHot Temperatureen_US
dc.subject.meshHumansen_US
dc.subject.meshImpetigo --microbiologyen_US
dc.subject.meshPharyngitis --microbiologyen_US
dc.subject.meshPolymerase Chain Reaction --methodsen_US
dc.subject.meshSequence Analysis, DNAen_US
dc.subject.meshStreptococcal Infections --microbiologyen_US
dc.subject.meshStreptococcus pyogenes --classificationen_US
dc.titleEvaluation of simplified DNA extraction methods for emm typing of group A streptococci.en_US
dc.typeEvaluation Studiesen_US
dc.typeJournal Articleen_US
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