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Evaluation of simplified DNA extraction methods for emm typing of group A streptococci.
Indian J Med Microbiol ; 2006 Apr; 24(2): 127-30
Article in English | IMSEAR | ID: sea-53445
ABSTRACT
Simplified 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.
Subject(s)
Full text: Available Index: IMSEAR (South-East Asia) Main subject: Streptococcal Infections / Streptococcus pyogenes / Bacterial Outer Membrane Proteins / Bacterial Proteins / Humans / DNA, Bacterial / Pharyngitis / Child / Child, Preschool / Polymerase Chain Reaction Type of study: Evaluation studies / Prognostic study Language: English Journal: Indian J Med Microbiol Journal subject: Microbiology Year: 2006 Type: Article

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Full text: Available Index: IMSEAR (South-East Asia) Main subject: Streptococcal Infections / Streptococcus pyogenes / Bacterial Outer Membrane Proteins / Bacterial Proteins / Humans / DNA, Bacterial / Pharyngitis / Child / Child, Preschool / Polymerase Chain Reaction Type of study: Evaluation studies / Prognostic study Language: English Journal: Indian J Med Microbiol Journal subject: Microbiology Year: 2006 Type: Article