DETERMINATION OF THE SEROTYPE PREVALENCE OF Streptococcus pneumoniae SEROGROUP 6 IN INDONESIA BY USING BsmAI RESTRICTION ENZYME

• Main Author: Monica Bernadette, Felicia
• Format: Final Project
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/44703
• Institution: Institut Teknologi Bandung

Streptococcus pneumoniae serogrup 6, consists of serotype 6A, 6B, 6C, 6D, 6E, 6F, and 6G, is one of the major serogroup causing invasive pneumococcal disease (IPD) worldwide. Several studies showed that serogroup 6 was the dominant serogroup in Indonesia. However, until now, there is no study about the serotype prevalence of S. pneumoniae serogroup 6 in Indonesia although the data is needed in the implementation of national pneumococcal vaccination. Determination of serotype (serotyping) for serogroup 6 is usually done by the combination of PCR and sequencing method, thus it is not efficient. Therefore, this study aimed to develop an alternative serotyping method for S. pneumoniae serogroup 6 so that it can be used to determine the serogroup 6 prevalence in Indonesia. In this study, we used 82 archived isolates of S. pneumoniae serogroup 6 belonged to Eijkman Institute. Serotyping was performed by PCR method to separate serotype 6A/6B from 6C/6D. Then, each serotype was separated by BsmAI restriction enzyme (recognition site: 5’ GTCTCN 3’) that will specifically cut the wciP of serotype 6B and 6D. Restriction results were confirmed by sequencing method. Furthermore, PCR method was used to differentiate serotype 6E from 6A and serotype 6F/6G from 6C and 6D. In this research, all results obtained by BsmAI were in concordance with those obtained by sequencing. Restriction of wciP genes of serotype 6B and 6D resulted in 120 bp band, whereas restriction didn’t occur on all serotypes 6A and 6C. The combination of PCR and restriction succeeded in accurately identifying 37 isolates as serotype 6A and 6B, 7 isolates serotype 6C, and 1 isolate serotype 6E. But, we didn’t find any 6D, 6F, and 6G isolates. In conclusion, serotype 6A and 6B were the dominant serotypes among S. pneumoniae serogroup 6 in Indonesia and BsmAI can be used effectively and efficiently for serotyping S. pneumoniae serogrup 6