Tesis

Karakterisasi Molekuler Streptococcus pneumoniae Resistan Multi-Obat Serotipe 19F dan 19A di Indonesia: Pilus, Gen Resistansi, Biofilm dan Tipe Sekuen = Molecular Characterization of Multidrug Resistant Streptococcus pneumoniae Serotype 19F and 19A in Indonesia: Pilus, Gene Resistance, Biofilm, and Sequence Type.

Streptococcus pneumoniae (S. pneumoniae) merupakan penyebab pneumonia, bakterimia dan meningitis. Penggunaan antibiotik yang kurang bijak menyebabkan berkembangnya klon resistan S. pneumoniae dan membuat kegagalan pengobatan infeksi S. pneumoniae. Pilus dan biofilm merupakan faktor virulensi yang penting pada S. pneumoniae. Strain S. pneumoniae resistan multi-obat di Indonesia didominasi oleh serotipe 19F dan 19A. Namun penelitian karakterisasi genomik molekuler dari strain tersebut belum pernah dilaporkan di Indonesia. Penelitian ini bertujuan untuk menganalisis gen resistansi, pilus, kemampuan pembentukan biofilm, dan tipe sekuen pada 104 isolat resistan multi-obat serotipe 19F dan 19A di Indonesia. PCR konvensional digunakan untuk mendeteksi gen resistansi terhadap makrolida (ermB dan mefA), tetrasiklin (tetM), dan kotrimoksazol (folA dan folP). Sekuensing DNA dilakukan untuk mengetahui mutasi yang menyebabkan resistansi kotrimoksazol. Sedangkan multilocus sequence typing (MLST) dikerjakan untuk mengidentifikasi tipe sekuen. Gen mefA, ermB, dan tetM ditemukan pada 93%, 52%, dan 100% isolat MDRSP. Insersi arginin dan prolin pada asam amino ke-59 dan Ile-100-Leu merupakan mutasi yang paling banyak ditemukan pada gen folP dan folA. Pilus tipe 1 dan tipe 2 ditemukan pada 93% dan 82% isolat resistan multi-obat. Kebanyakan isolat resistan multi-obat tidak dapat membentuk biofilm (64.42%). Sedangkan proporsi tipe sekuen dari 10 isolat yang diuji adalah ST1464 (50%), ST320 (20%), ST271 (10%), ST236 (10%), dan ST5047 (50%). Hasil studi ini menggambarkan virulensi strain resistan multi-obat serotipe 19F dan 19A di Indonesia. Hasil penelitian ini dapat digunakan untuk pengembangan pengobatan infeksi yang dikarenakan strain S. pneumoniae resistan multi-obat, serta mengontrol dan memonitor penyebaran klon resistan dan virulen di Indonesia.
Kata Kunci: Streptococcus pneumoniae resistan multi-obat, gen resistansi, pilus, biofilm, tipe sekuen


Streptococcus pneumoniae (S. pneumoniae) is a major cause of bacterial pneumonia, bacteremia, and meningitis. Inappropriate use of antibiotics has led to the development of resistant pneumococcal clones and the failure of pneumococcal infections treatment. Meanwhile, pilus and biofilm are important S. pneumoniae virulence factors. Multidrug resistant S. pneumoniae (MDRSP) in Indonesia were dominated by serotype 19F and 19A. However, molecular genomics study of these strains has not been reported. This study aims to analyze resistance genes, pilus, biofilm formation ability, and sequence type (ST) among 104 isolates of MDRSP serotypes 19F and 19A in Indonesia. Conventional PCR was used to detect resistance genes, namely macrolide (ermB and mefA), tetracycline (tetM), and cotrimoxazole (folA and folP) gene. DNA sequencing was performed to investigate mutations in genes folA and folP. Multilocus sequence typing (MLST) was conducted to identify ST. The mefA, ermB, and tetM genes were found in 93%, 52%, and 100% MDRSP, respectively. The insertion of arginine and proline, and Ile-100-Leu were the most common mutations in the folP and folA. Pilus type 1 and type 2 were discovered in 93% and 82% of MDRSP isolates. The majority of isolates were not able to form biofilm (64%). While the ST proportion of the 10 isolates tested were ST1464 (50%), ST320 (20%), ST271 (10%), ST236 (10%), and ST5047 (50%). This study showed the virulence of MDRSP strains serotypes 19F and 19A in Indonesia. The results of this study could be used to develop treatment S. pneumoniae infection caused by multi-drug resistant strains, as well as to control and monitor the spread of resistant and virulent clones in Indonesia.
Keywords: Multidrug resistant Streptococcus pneumoniae (MDRSP), gene resistance, pilus, biofilm, and sequence type