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Evaluation of Modified Sequential Multiplex PCR for Streptococcus pneumoniae Serotyping.

The aims of this study were to develop modified SM-PCR primer sets and to evaluate their ability and efficiency for serotype determination. We selected target serotypes for SM-PCR testing according to the prevalence of serotypes as reported in papers published in Asian countries. The modified SM-PCR consisted of six groups of PCRs, with each reaction being carried out using five pairs of primers. We evaluated the efficiency and performance of this modified multiplex PCR using 378 pneumococcal strains by comparing the findings with the results of the Quellung reaction. A total of 30 primer pairs were used for a consecutive set of six reactions. All results were concordant with those of the Quellung reaction, and there was no cross reactivity on multiplex PCR. We could identify the final serotypes of 370 isolates (97.9%). The coverage rates of modified SM-PCR were 42.6%, 65.8%, and 79.4% in Reactions 1, 2, and 3, respectively. The modified SM-PCR showed good performance in detecting pneumococcal serotypes and could be used as a helpful alternative to the Quellung reaction.

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