WIWIK PURWANTI, 091214353005 (2014) IDENTIFIKASI GENOTYPE Salmonella typhi DARI ISOLAT BEBERAPA DAERAH DI INDONESIA BERDASARKAN FILOGENI. Thesis thesis, UNIVERSITAS AIRLANGGA.

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Abstract

The purpose of this study was to identify the genotype of isolates of Salmonella typhi from several regions in Indonesia based phylogeny to determine the pattern of spread of strains. Basic mapping genotypes of Salmonella typhi in Indonesia is to reveal the diversity of strains S.typhi isolates originating from several regions in Indonesia. This research is a descriptive observational study of molecular epidemiology approach. The design used a cross-sectional study. The samples were isolates of S.typhi from several regions in Indonesia. Large sample is determined by 15 isolates. All isolates from these regions after a confirmed positive with microbiology then isolate DNA extraction with Presto™ gDNA Mini Bacteria Kit device subsequent DNA amplification using specific primers with 3 pairs of green taq mix PCR kit with loding dye. PCR products were visualized on agarose gel, purified, labeled and in sequencing. S.typhi genotypes in 15 isolates from several regions in Indonesia were identified based on 16S rRNA gene sequences. Base sequence of the 16S rRNA gene sequences were analyzed together with sequences of 16S rRNA and representatives of the family Enterobacteriaceae S.typhi is E.coli obtained from the NCBI database using the program Clustal W. Phylogeny tree constructed using Mega 6.0 program while nucleotide homology and differences were analyzed with the program Bioedit. The results showed that there were similarities among the isolates S.typhi ranged from 98.0 to 99.6%. Phylogeny analysis showed that all isolates form a new diversity center with S.typhi 16S rRNA (Z47554). The result of sensitivity to antibiotics showed that 8 (53.3%) isolates were resistant to ampicillin S.typhi. Conclusion that there are 2 clade, although all isolates were included in the same clade genotypes with S.typhi but still no diversity in the group. The difference in this clade 2 can be used as a preliminary test for the development of diagnostic kits S.typhi.

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