Evaluation of simplified DNA extraction methods for emm typing of group A streptococci.
Indian J Med Microbiol
;
2006 Apr; 24(2): 127-30
Artigo
em Inglês
| 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.
Texto completo:
DisponíveL
Índice:
IMSEAR (Sudeste Asiático)
Assunto principal:
Infecções Estreptocócicas
/
Streptococcus pyogenes
/
Proteínas da Membrana Bacteriana Externa
/
Proteínas de Bactérias
/
Humanos
/
DNA Bacteriano
/
Faringite
/
Criança
/
Pré-Escolar
/
Reação em Cadeia da Polimerase
Tipo de estudo:
Estudos de avaliação
/
Estudo prognóstico
Idioma:
Inglês
Revista:
Indian J Med Microbiol
Assunto da revista:
Microbiology
Ano de publicação:
2006
Tipo de documento:
Artigo
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