Molecular determinants of artemisinin resistance in k13 gene of plasmodium falciparum

Artemisinin-based combination therapy (ACT) is the first-line therapy in most malaria endemic countries. An impressive 47% reduction in the global mortality rate between 2000 and 2013 has been achieved by ACT and artemisinin (ART) monotherapy. However, artemisinin resistance (AR) by Plasmodium falciparum (P. falciparum) is now prevalent across south-east Asia (SEA). AR is indicated by delayed parasite clearance of more than 3 days after standard ART treatment and reduced in vitrosusceptibility. Recent work has shown association of AR with mutations in the propeller domain of the kelch gene on chromosome 13 (PF3D7_1343700, k13gene) of P. falciparum. The C580Y mutation of thek13gene is highly prevalent in Cambodia, Myanmar and eastern and western Thailand, while the F446I mutation is predominant in the China-Myanmar border regions as well as in Myanmar. AR has not reached India and Africa, where non-synonymous mutations not associated with delayed parasite clearance are present. Because the location of Myanmar is central between SEA and Africa, a country-specific strategy for Myanmar Artemisinin Resistance Containment (MARC) is necessary. Moreover, regular periodic tracking of prevalent molecular determinants such as C580Y and F446I mutations will be beneficial

Evaluation of PAIusp subtyping to characterize uropathogenic E. coli isolates

Introduction: Uropathogenic virulence factors have been identified by comparing the prevalence of these among urinary tract isolates and environmental strains. The uropathogenic-specific protein (USP) gene is present on the pathogenicity island (PAI) of uropathogenic Escherichia coli (UPEC) and, depending on its two diverse gene types and the sequential patterns of three open reading frame units (orfUs) following it, there is a method to characterize UPEC epidemiologically called PAIusp subtyping. Methodology: A total of 162 UPEC isolates from Sabah, Malaysia, were tested for the presence of the usp gene and the sequential patterns of three orfUs following it using polymerase chain reaction (PCR). In addition, by means of triplex PCR, the prevalence of the usp gene was compared with other two VFs of UPEC, namely alpha hemolysin (α-hly) and cytotoxic necrotizing factor (cnf-1) genes encoding two toxins. Results: The results showed that the usp gene was found in 78.40% of UPEC isolates, indicating that its prevalence was comparable to that found in a previous study in Japan. The two or three orfUs were also associated with the usp gene in this study. All the PAIusp subtypes observed in Japan were present in this study, while subtype IIa was the most common in both studies. The usp gene was observed in a higher percentage of isolates when compared with ɑ-hly and cnf-1 genes. Conclusions: The findings in Japan and Sabah, East Malaysia, were similar, indicating that PAIusp subtyping is applicable to the characterization of UPEC strains epidemiologically elsewhere in the world

Detection of sequence type 131 in multi-drug resistant uropathogenic Escherichia coli isolates from two hospitals of Sabah

Background: Escherichia coli sequence type 131 (ST131) has emerged among bacteria causing urinary tract infection (UTI) in the previous decade. This ST contains multiple drug resistant (MDR) genes together with genes encoding many virulence factors. As a result, this strain of uropathogenic E. coli (UPEC) gives rise to treatment failure with consequent prolonged stay in a hospital. Therefore, earlier identification of this strain in the hospital has advantage in combating severe type of UTI. Objective: To detect ST 131 strains in MDR UPEC isolates from two hospitals of Sabah. Materials and Methods: Antibiotic susceptibility tests were performed to detect MDR isolates. Two polymerase chain reactions (PCRs) including mdh and gyrB allelic-specific PCR were performed on these MDR to detect ST131 strains. Results: The results showed four isolates were resistant to TMP-SMX, gentamycin, ciprofloxacin, and cefotaxime, and three isolates of these were investigated to be ST131 clones by two PCR reactions. Conclusion: There is the presence of ST131 strains in hospitals of Sabah. This information will be a guideline for the clinician in the management of UTI in the clinical settings

Distribution of sul genes and their variants in uropathogenic Escherichia coli isolated from two hospitals of Sabah

Sulphonamides resistant strains are highly prevalent in uropathogenic Escherichia coli (UPEC) isolates. Sul genes encode sulphonamide resistance and are present on transferrable plasmids. Integrons (IGNs) are genetic elements containing integrase gene, attl site and gene cassettes which carry multiple antibiotic resistant genes. Class 1 integrons have been extensively studied because these were most prevalent among clinical isolates. In this study, UPEC isolates were determined for the antibiotic susceptibility patterns to four antibiotics commonly used for urinary tract infections, which include co-trimethoxazole (TMP-STX). Distribution of sul genes and integrase1 gene (intI1) was studied in TMP-STX resistant UPEC isolates by using multiplex polymerase chain reaction (mPCR). Sul genes variants were investigated by DNA sequencing of the whole open reading frame of sul1 and sul2 genes and PCR product of sul3 gene. Sul1, sul2 and sul3genes were prevalent in 37 (24.7%) of 150 UPEC isolates. IntI1 is positive in 22 sul genes positive isolates. Of six isolates positive with sul2 genes, sul2(a) and sul2(b) variants, which were described in the previous study, in the four isolates and the two isolates respectively were observed. This is the first mPCR which investigates the prevalence of three sul genes and intI1 in the UPEC clinical isolates from two hospitals of Sabah

Genetic diversity of toxigenic Vibrio cholerae O1 from Sabah, Malaysia 2015

Background Cholera is an important health problem in Sabah, a Malaysian state in northern Borneo; however, Vibrio cholerae in Sabah have never been characterized. Since 2002, serogroup O1 strains having the traits of both classical and El Tor biotype, designated as atypical El Tor biotype, have been increasingly reported as the cause of cholera worldwide. These variants are believed to produce clinically more severe disease like classical strains. Purpose The purpose of this study is to investigate the genetic diversity of V. cholerae in Sabah and whether V. cholerae in Sabah belong to atypical El Tor biotype. Methods ERIC-PCR, a DNA fingerprinting method for bacterial pathogens based on the enterobacterial repetitive intergenic consensus sequence, was used to study the genetic diversity of 65 clinical V. cholerae O1 isolates from 3 districts (Kudat, Beluran, Sandakan) in Sabah and one environmental isolate from coastal sea water in Kudat district. In addition, we studied the biotype-specific genetic traits in these isolates to establish their biotype. Results Different fingerprint patterns were seen in isolates from these three districts but one of the patterns was seen in more than one district. Clinical isolates and environmental isolate have different patterns. In addition, Sabah isolates harbor genetic traits specific to both classical biotype (ctxB-1, rstRCla) and El Tor biotype (rstRET, rstC, tcpAET, rtxC, VC2346). Conclusion This study revealed that V. cholerae in Sabah were genetically diverse and were atypical El Tor strains. Fingerprint patterns of these isolates will be useful in tracing the origin of this pathogen in the future

Molecular characterization of Mycobacterium species isolates from patients with pulmonary tuberculosis in Sabah, Malaysia

Tuberculosis (TB) is one of the deadliest diseases worldwide, caused by members of Mycobacterium tuberculosis complex (MTBC), commonly by Mycobacterium tuberculosis (Mtb) and Mycobacterium bovis. In Malaysia, Sabah is one of the states of public health concern with the highest TB cases. Clinical presentations of TB and non-tuberculous mycobacteria (NTM) lung disease are similar, and mycobacteria appear to be identical under standard diagnosis with sputum smear microscopy, causing difficulty to diagnose TB. Identification of Mycobacterium species is essential for effective management of mycobacterial diseases treatment and their control strategy. Thus, this study aimed to identify the Mycobacterium species from suspected TB patients in Sabah using molecular methods. Sputum samples (n=595) were screened with GeneXpert MTB/RIF (Xpert), and positive TB samples (n=67) were processed and cultured in BACTEC MGIT. Forty-five isolates were successfully recovered in MGIT and characterisation of the mycobacterial isolates using PCR and/or sequencing with rpoB, RD9, hsp65, and 16S rRNA genes confirmed the presence of Mtb in 41 samples, and four non-mycobacteria, i.e. Microbacterium laevaniformans, Streptomyces sp., Streptomyces misionensis and Gordonia sp. These non-mycobacteria isolates showed negative results when tested directly with Xpert. In conclusion, Mtb is the predominant species of MTBC circulating in Sabah. The presence of non-mycobacteria in this study was due to bacterial contamination in MGIT, not bacterial cross-reactivity in Xpert, implying the high sensitivity and specificity of Xpert for diagnosis of TB

Molecular characterization of mycobacterium species isolates from patients with pulmonary tuberculosis in Sabah, Malaysia

Tuberculosis (TB) is one of the deadliest diseases worldwide, caused by members of Mycobacterium tuberculosis complex (MTBC), commonly by Mycobacterium tuberculosis (Mtb) and Mycobacterium bovis. In Malaysia, Sabah is one of the states of public health concern with the highest TB cases. Clinical presentations of TB and non-tuberculous mycobacteria (NTM) lung disease are similar, and mycobacteria appear to be identical under standard diagnosis with sputum smear microscopy, causing difficulty to diagnose TB. Identification of Mycobacterium species is essential for effective management of mycobacterial diseases treatment and their control strategy. Thus, this study aimed to identify the Mycobacterium species from suspected TB patients in Sabah using molecular methods. Sputum samples (n=595) were screened with GeneXpert MTB/RIF (Xpert), and positive TB samples (n=67) were processed and cultured in BACTEC MGIT. Forty-five isolates were successfully recovered in MGIT and characterisation of the mycobacterial isolates using PCR and/or sequencing with rpoB, RD9, hsp65, and 16S rRNA genes confirmed the presence of Mtb in 41 samples, and four non-mycobacteria, i.e. Microbacterium laevaniformans, Streptomyces sp., Streptomyces misionensis and Gordonia sp. These non-mycobacteria isolates showed negative results when tested directly with Xpert. In conclusion, Mtb is the predominant species of MTBC circulating in Sabah. The presence of non-mycobacteria in this study was due to bacterial contamination in MGIT, not bacterial cross-reactivity in Xpert, implying the high sensitivity and specificity of Xpert for diagnosis of TB

Health promotion activity in Kampung Rosob, Pitas

We, group 3 members which consist of 9 medical students, 8 who recently finished 1st year Medical program and 1 who recently finished 2nd year Medical program and 5 nursing students went to Kg Rosob, Pitas for health promotion from 24th August 2015 to 2nd September 2015. During our visit in the village, we started with opening ceremony once we reached the village as we have already planned with the head of the village to do opening ceremony before any activities in the village. The next day, we started our rapid rural appraisal (RRA) to get overview of the village and health problems and then we planned activities according to information from RRA to promote the health of villagers by various actions such as 1. RRA which indudes direct or field observation, secondary data resources, key informant Interview, individual interview, group meeting or interview and discussion, house-house visit, and social mapping. 2. Health education by means of talk such as breast and cervix cancer , demonstration , exhibitions , and health screening . 3. Life style and behavioral changes; for example: talk for high blood pressure , personal hygiene practice, school activities ,mini sports day and aerobic exercise. 4. Nutritional education; for example: talk on balanced diet, proper way to take medicine and good eating habits. 5. Education ; fun learning for year 5 students as a preparation for them to sit for next year UPSR examination. And involved in motivation camp for year 6 students as a preparation for their UPSR examination in August. 6.Voluntary works ; such as gotong royong to clean the compound of the village for the benefits of their health, and also killing the hair louse for the infected girls in the dorm