Detection and characterization of Uropathogenic Escherichia Coli Sequence type 131 in Sabah, Malaysia

• Main Author: Lai, Yun Mei
• Format: Thesis
• Language: English; English
• Published: 2018
• Subjects: RA Public aspects of medicine
• Online Access: https://eprints.ums.edu.my/id/eprint/26955/1/Detection%20and%20characterization%20of%20Uropathogenic%20Escherichia%20Coli%20Sequence%20type%20131%20in%20Sabah.pdf; https://eprints.ums.edu.my/id/eprint/26955/2/Detection%20and%20characterization%20of%20Uropathogenic%20Escherichia%20Coli%20Sequence%20type%20131%20in%20Sabah%2C%20Malaysia%2024%20pages.pdf; https://eprints.ums.edu.my/id/eprint/26955/

Escherichia coli (E. coli) Sequence type 131 (ST131) is the multi-drug resistant pathogen causing urinary tract infection (UTI) as well as other extra-intestinal infections and intestinal infections in human. It is the pandemic pathogen and spreading worldwide. Mega-plasmids carried by ST131 contain multiple drug resistance gene and virulence encoding genes together. MLST was used to identify this pathogen by sequencing seven housekeeping genes for each sample. The heavy workload and time-consuming procedure of MLST ST131 need to be overcome for the detection of ST131. In Sabah, the presence of ST131, ST131 clade C sub-linages (ST131H30, ST131H30R and ST131H30Rx), drug resistance genes profile, the virulence factor profile and type of replicon in ST131 remained unknown. In this study, Uropathogenic E. coli (UPEC) 294 samples were isolated from UTI patients from Hospital Queen Elizabeth I and Hospital Wanita Dan Kanak-Kanak Sabah. From these samples, multi-drug resistant isolate was detected by antibiotic susceptibility tests. ST131 isolates were investigated among these ciprofloxacin resistant samples by Achtman Scheme. Duplex mismatch amplification mutation assay - Polymerase Chain Reaction (MAMA-PCR) was developed with the identified ST131 isolates. Clade C sub-lineage was identified among the ST131 isolates. Common virulence encoding genes, drug resistance genes and types of replicon were identified from the ST131 isolates. Seventeen ST131 isolates were identified among the UPEC isolates. Duplex MAMA-PCR was applied as the rapid detection method for detection of ST131. Among the ST131 isolates, nine isolates were ST131H30Rx sub-lineage. Only one ST131 isolate carried eight common drug resistance genes. In this study, usp, hlyA and cnf-1 were absent in ST131 isolates. OmpT, fyuA, iutA, kpsmII and fimH were 100% present among the ST131 isolates. F1B and Frep were the common replicon types found among the ST131 isolates. This is the pioneer study on ST131 isolates in Malaysia as well as in South East Asia region. This study showed ST131 are frequently presence in gentamicin resistant UPEC isolates rather than ciprofloxacin resistant UPEC isolates. Different drug resistance gene profile and virulence factor profile showed the unique characteristics of ST131 isolates. Replicon types F1B and Frep were common and co-exist, a finding which was similar to the previous study. Study showed the presence of ST131 in the collected isolates and ST131H30Rx was the common sub-lineage. Duplex MAMA-PCR was successfully developed as the rapidmethod for ST131 detection. Molecular characteristics observed in this study will provide important information for the future study.