Introduction: Tuberculosis (TB), commonly caused by Mycobacterium tuberculosis (Mtb), is one of the ten leading causes of death worldwide. The gold standard, microbiological culture for detection and differentiation of mycobac-teria are time-consuming and laborious. The use of fast, easy and sensitive nucleic acid amplification tests (NAATs) for diagnosis of TB remains challenging because there is a high degree of homology within Mtb complex (MTBC) members and absence of target genes in the genome of some strains. This study aimed to identify new candidate genetic marker and to design specific primers to detect Mtb using in silico methods. Methods: Using Basic Local Alignment Search Tool (BLAST) program, Mtb H37Rv chromosome reference genome sequence was mapped with other MTBC members and a single nucleotide polymorphism (SNP) at Rv1970 was found to be specific only for Mtb strains. Mismatch amplification mutation assay (MAMA) combine with polymerase chain reaction (PCR) was used as an alternative method to detect the point mutation. MAMA primers targeting the SNP were designed using Primer-BLAST and the PCR assay was optimized via Taguchi method. Results: The assay amplified a 112 bp gene fragment and was able to detect all Mtb strains, but not the other MTBC members and non-tuberculous Mycobacte-ria. The detection limit of the assay was 60 pg/μl. Conclusion: Bioinformatics has provided predictive identification of many new target markers. The designed primers were found to be highly specific at single-gene target resolution for detection of Mtb.
Prenatal exposure to chronic stress during critical periods of foetal development produces depression, attention and learning deficits, hormonal imbalances and affects the brain. The effect of prenatal restraint-stress on the postnatal developmental milestones, anthropometric measurements, and the body, brain and adrenal gland weights of the pups were examined and compared with the unrestrained control and the restrained group under the pyramid at postnatal day 10 and 21. Pregnant rats were restrained (9h/day) from gestation day 7 until parturition. Results showed significant delay in the milestones by one day in the restraint control (RC) compared to the unrestrained normal control (NC), while pups of the restrained pyramid (RP) group did not show the delay. Significant decreases in the anthropometric measurements, body and brain weights in RC group were observed at both postnatal days, while the RP group results matched with the NC group. Significant increase in the adrenal weights was found in the RC group compared to NC group and not the RP group. Results suggest prenatal restraint-stress definitely hampers the developmental milestones, anthropometric measurements, and body and brain weights of the young offspring. Results suggest, pyramid environment counteracts and protects the deleterious effects of chronic prenatal stress.
Introduction:Dermatoglyphics have been used extensively to characterize populations, analyse the nature and origin of human variation and population structure and evaluate the micro-differentiation of populations. Dermato-glyphics is considered as a sensitive indicator of intrauterine anomalies and known to be one of the best available diagnostic tools in genetic disorders. KadazanDusun and Bajau are the major ethnic groups of Sabah. The aim of the study was to study the distribution of fingerprint patterns on different digits in these two ethnicities of Sabah and to find if any variation occurs in fingerprint pattern qualitatively between these ethnicities. Methods: This cross-section-al study was conducted from March’ 2016 to February’ 2018 at different villages of Kudat, Ranau, Kota Belud, Kota Kinabalu and Papar, Sabah. After obtaining informed consent 400 persons (KadazanDusun 250, Bajau 150) were included in this study by stratified random sampling. Demographic data and history of present and past illness were obtained from each subject through a valid questionnaire. Finger and palm prints were photographed using Nikon D5200 camera and later were grouped into different types manually. Results: There were 205 female and 195 male participants in the study. Among the three types of fingerprints, whorls were more abundant (>50%) in the thumbs, index, ring and little fingers whereas loops were observed more (>55%) in the middle fingers. The arches were the least (0-3%) observed pattern in all the fingers. The dermatoglyphic pattern of right and left index fingers were only significantly different (p0.05) difference in respect of left and right hands and gender. Conclusion: The study findings would serve as a baseline data for further studies to develop data base of the dermatoglyphic pattern of major ethnic groups of Sabah which would facilitate non-invasive diagnosis of congenital anomalies and various genetic disorders.
Assessment method of medical students by conducting examination is to identify the quality and quantity of their academic performance. Essay paper is one of the most common assessment tools in the Faculty of Medicine and Health Sciences, Universiti Malaysia Sabah. Double marking is a means by which academic staff attempts to produce fair results for the students. Eighty eight medical students sat for three sets of Essay Papers of Professional I examination in March 2012. The double marking on the essay papers was done by two lecturers of each clinical department concerned for each discipline. Inter-examiner agreement and its effect on the reliability of the final score for the students were calculated by using Kappa statistics and Intra-class Correlation Coefficient (ICC). Reliability coefficient of the scores were also calculated for the different disciplines. In Part A essay paper, Cohan’s Kappa was 0.48 (p
Emergence of multidrug resistant tuberculosis (MDR-TB) and extensively drug resistant tuberculosis (XDR-TB) is one of the reasons why tuberculosis (TB) continues to cause great mortality and morbidity in less-developed countries. The development of rapid diagnostic methods targeting genetic mutations associated with resistance to the anti-tuberculous drugs is essential to fight this deadly pathogen. Isoniazid (INH) has been included in the multidrug regimens for the treatment of drug-susceptible TB for the decades. In the worldwide setting, isoniazid resistance was highly prevalent and was observed in one of every seven TB cases. Since katG315 mutation is highly prevalent, the common mutation in the enzyme essential for the activation of the INH concerned with the mechanism of drug resistance and associated with high level resistance to INH, katG315 mutation was necessary to be identified by molecular method as a molecular determinant of INH resistant Mycobacterium tuberculosis. The prevalence of katG315 mutation in various countries was discussed in this report and a new molecular method for the detection of the mutation was proposed.
Introduction: Tuberculosis (TB) still remains a public health problem worldwide and the emergence of drug resistant TB (DR-TB) has worsened the situation as it is difficult and expensive to treat. The characterization of the genetic mutations underlying streptomycin resistance may be helpful in developing rapid detection methods which may guide clinicians in making therapeutic decisions. The aim of this study is to detect mutations causing streptomycin (STR) resistance in Mycobacterium tuberculosis isolates from Sabah. Methods: Susceptibility testing was carried out in MGIT system for 42 Mycobacterium tuberculosis clinical isolates. The drug resistant isolates were subject to whole genome sequencing and in-silico analysis was performed to detect the mutations in the sequence of the rpsL gene known to confer resistance to anti-tuberculous drugs. Results: Of the 42 positive isolates, 27 (64.3%) are shown to be susceptible towards first line drugs (FLDs) while 15 (35.7%) isolates were mono- and multiple resistant to the FLDs. Our findings reveal that the isolate 145 possess mutations at codon 43 within rpsL gene with amino acid change A to G (K43R). Conclusion: Findings from this study enable us to expand our knowledge of mutations causing drug resistance in Mycobacterium tuberculosis and the point mutations, which can be used as the potential marker for detection of drug resistant isolates.