This study characterized carriage and clinical pneumococcal isolates for serotypes, penicillin susceptibility, virulence genes and restriction fragment length polymorphism (RFLP) pattern of penicillin binding protein (PBP) genes. DNA fingerprint of isolates was generated by BOX-PCR. Majority of serotypes were 23F followed by 19F, 19A and 6A. Twenty-four percent of isolates were penicillin non-susceptible (PNSP). All of the targeted virulence genes were detected in all isolates with the exception of pili; 20.6% (n=22) for PI-1 and 14.0% (n=15) for PI-2. Of the 13 isolates which carried both PI-1 and PI-2, 10 were of clinical origin. Digested pbp-DNA produced three PBP-RFLP profiles for pbp1a (A1 to A3), six profiles for pbp2b (B1 to B6) and seven for pbp2x (X1 to X7) mostly in PNSPs. Based on BOX-PCR analysis, the majority of isolates were genetically diverse with a small number of potentially related isolates carrying pili genes. No obvious genotypic association was observed pertaining to carriage and clinical origin of isolates.
Background: Cervical cancer is the third leading cause of death in Malaysia, and Human Papilloma Virus (HPV) is the principal aetiology that is responsible for its development. This study was aimed to determine the prevalence and distribution of HPV types among different age groups, ethnicity, and areas in Malaysia. Materials and Methods: A total of 764 women aged 20-74 years old within the cities of Johor Bahru, Kuala Lumpur, Ipoh, Penang, and Kota Kinabalu underwent both cervical cytological assessment and HPV DNA analysis. Cervical cytology glass slides were prepared using the liquid base technique (Path TEZT TM). HPV DNA was extracted using TANBead ® Nucleic Acid Extraction Kit (Taiwan Advanced Nonotech Inc.), then the types were further identified using a DR.HPV Genotyping IVD kit. Results: The prevalence of HPV infection was 14.0% (107/764) with high-risk type at 10.7% (82/764) and low-risk type at 3.27% (25/764). The most common high-risk HPV types were HPV-52, 66, 33, 39, and 58 whereas low-risk HPV types were HPV-6, 40, and 81. The majority of HPV infections (80.37%) were detected in women with normal cytology results. The most prevalent HPV type among Chinese is 33 (n=6) followed by 16, 44, 58, 66 and 68 (n=5). Among Malays, HPV 16 and 51 were the two most prevalent types (n=2). The sensitivity of the HPV DNA test compared to cytology was 100% with a specificity of 88.37%. Conclusion: This study revealed that the most common high-risk HPV type among women living in urban areas in Malaysia is HPV 52, unfortunately which is not the type of infection the current HPV vaccine is covered for protection among females. These findings may contribute beneficial information to health care providers for the appropriate use of HPV vaccine in the prevention of cervical cancer in Malaysia.
The global emergence of methicillin-resistant Staphylococcus aureus (MRSA) that unsusceptible to a wide selection of antimicrobial agents and any newly introduced antimicrobial over the past decades has triggered more extensive holistic measures to put an end to this situation. Molecular surveillance of MRSA clones is important to understand their evolutionary dynamics for investigating outbreaks, propagating precautionary measures, as well as planning for appropriate treatment. This review includes peer-reviewed reports on the molecular characterisation of clinical Staphylococcus aureus isolates within Malaysian hospitals from year 2008 to 2020. This work highlights the molecular clones of hospital-acquired MRSA (HA-MRSA) and community-acquired MRSA (CA-MRSA) isolates from Malaysian hospitals, with description on their ever-changing pattern. Among HA-MRSA, the ST22-t032-SCCmec IV MRSA clone was reported to supplant the previous dominating clone, ST239-t037-SCCmec III. Meanwhile, ST30, ST772, ST6 and ST22 were repeatedly detected in CA-MRSA, however, none of the strains became predominant. Future in-depth study on molecular epidemiology of MRSA clone is essential for the investigation of the extent of the clonal shift, especially in Malaysia.
Aims: To develop a real-time polymerase chain reaction system Vi-qPCR in the detection of Salmonella enterica serovar Typhi (S. Typhi), targeting the vexC gene encoding for Vi antigen (capsular polysaccharide antigen) and to evaluate its sensitivity and specificity performance using pure cultures of S. Typhi and other enteric pathogens. Methodology and results: Microbiological, biochemical and serotyping tests were conducted to determine the phenotypic characteristics of S. Typhi and other enteric pathogens in our collection. Primers were designed using Primer3 software and their in-silico specificity were analysed using Basic Local Alignment System Tool (BLAST). Optimisation of PCR annealing temperature was done prior to assessment of sensitivity and specificity performance against artificial serially diluted seeded stools. The primers were found to be 100% specific in the detection of S. Typhi towards 32 tested clinical strains. Verification of gene amplification by comparing the nucleotide sequences against reference genes in the GenBank database revealed high specificity to S. Typhi. Statistical analysis indicates that this method results in 100% sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV). Moreover, Vi-qPCR allows the detection of S. Typhi as low as 131.4 CFU/g of stool sample. Conclusion, significance and impact of study: A rapid and sensitive method for detection of Salmonella enterica serovar Typhi (S. Typhi) is desired as a diagnostic tool to improve typhoid management. The Vi-qPCR represent a promising non-invasive diagnostic tool for medical microbiology laboratories as a method for the detection of S. Typhi in both pure culture and stool specimens especially in chronic asymptomatic carriers where shedding of S. Typhi is intermittent and sometimes occurs in low level.
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