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Antifungal susceptibility of molecularly confirmed Aspergillus species from clinical samples

Tzar MN, Mustakim S, Yusoff H, Tap RM

Malays J Pathol, 2024 Apr;46(1):71-78.
PMID: 38682846

Invasive aspergillosis is the second most common invasive human mycosis but susceptibility data of Aspergillus species is limited. Antifungal treatment of aspergillosis is often done empirically without knowing the true susceptibility. Therefore, we aimed to determine antifungal susceptibility of Aspergillus species isolated from various clinical specimens over a 1-year period. We identified 28 Aspergillus isolates by sequencing the internal transcribed spacer (ITS) and β-tubulin genes and performed antifungal susceptibility testing on these isolates using Sensititre YeastOne. The isolates were identified as Aspergillus niger (60.7%), A. fumigatus (21.4%), A. flavus (10.7%), A. chevalieri (3.6%) and A. tubingensis (3.6%). Based on the European Committee on Antimicrobial Susceptibility Testing (EUCAST) Antifungal Clinical Breakpoint for Aspergillus spp., 16/17 (94.1%) A. niger isolates were susceptible to amphotericin B, all six isolates (100%) of A. fumigatus were susceptible to amphotericin B, itraconazole and voriconazole, but only 5/6 (83.3%) A. fumigatus were susceptible to posaconazole. Meanwhile, all three (100%) A. flavus isolates were susceptible to itraconazole. There are no other breakpoints established by the EUCAST for other antifungal-species combinations. In conclusions, Aspergillus niger remains the most commonly isolated species from clinical specimens and Aspergillus isolates at our centre are still largely susceptible to amphotericin B, echinocandins and most azoles. This information is valuable in guiding antifungal therapy in the treatment of aspergillosis.
Fulltext Neonatal malaria presenting as neonatal sepsis: a case report

Lim KY, Ang EL, Tan KK, Mustakim S

Malaysian Journal of Paediatrics and Child Health, 2016;22(1):48-57.
MyJurnal

Neonatal malaria may be overlooked likely due to its non-specific features and low prevalence in Malaysia. In this case report, we detail a case of neonatal malaria in an 18-day old baby girl of Myanmar origin who presented with 6 days of intermittent fever but was otherwise well. Initially, she was treated as neonatal sepsis. She then developed thrombocytopaenia and severe anaemia with persistent spikes of temperature. This prompted a series of investigations and multiple changes of antibiotics. The diagnosis of neonatal malaria surfaced when her peripheral blood film incidentally revealed the presence of Plasmodium vivax parasites. Peripheral blood smears are simple and inexpensive. Therefore practising especially in endemic areas for malaria, we need to consider this diagnosis when dealing with neonatal sepsis that does not respond to standard treatment.
The reliability of a rapid molecular detection method in determining the prevalence of rifampicin-resistant Mycobacterium tuberculosis in an urban district health facility in Malaysia

Ding CH, Ismail Z, Sulong A, Wahab AA, Gan B, Mustakim S, et al.

Malays J Pathol, 2020 Dec;42(3):401-407.
PMID: 33361721

INTRODUCTION: Rifampicin is a key first-line antimycobacterial agent employed for the treatment of pulmonary tuberculosis (PTB). This study sought to obtain prevalence data on rifampicin-resistant Mycobacterium tuberculosis among smear-positive PTB patients in the Klang District of Malaysia.
MATERIALS AND METHODS: A total of 103 patients from the Chest Clinic of Hospital Tengku Ampuan Rahimah with sputum smears positive for acid-fast bacilli were included in this cross-sectional study. All sputa were tested using Xpert MTB/RIF to confirm the presence of M. tuberculosis complex and detect rifampicin resistance. Sputa were also sent to a respiratory medicine institute for mycobacterial culture. Positive cultures were then submitted to a reference laboratory, where isolates identified as M. tuberculosis complex underwent drug susceptibility testing (DST).
RESULTS: A total of 58 (56.3%) patients were newly diagnosed and 45 (43.7%) patients were previously treated. Xpert MTB/RIF was able to detect rifampicin resistance with a sensitivity and specificity of 87.5% and 98.9%, respectively. Assuming that a single resistant result from Xpert MTB/RIF or any DST method was sufficient to denote resistance, a total of 8/103 patients had rifampicinresistant M. tuberculosis. All eight patients were previously treated for PTB (p<0.05). The overall prevalence of rifampicin resistance among smear-positive PTB patients was 7.8%, although it was 17.8% among the previously treated ones.
CONCLUSION: The local prevalence of rifampicin-resistant M. tuberculosis was particularly high among previously treated patients. Xpert MTB/RIF can be employed in urban district health facilities not only to diagnose PTB in smear-positive patients, but also to detect rifampicin resistance with good sensitivity and specificity.
A seven-year retrospective analysis of the clinicopathological and mycological manifestations of fungal rhinosinusitis in a single-centre tropical climate hospital

Goh LC, Shakri ED, Ong HY, Mustakim S, Shaariyah MM, Ng WSJ, et al.

J Laryngol Otol, 2017 Sep;131(9):813-816.
PMID: 28841131 DOI: 10.1017/S0022215117001505

OBJECTIVE: To evaluate the clinicopathological and mycological manifestations of fungal rhinosinusitis occurring in the Tengku Ampuan Rahimah Hospital, in Klang, Malaysia, which has a tropical climate.
METHODS: Records of patients treated from 2009 to 2016 were analysed retrospectively. Data from the records were indexed based on age, gender, clinical presentations, symptom duration, clinical signs and mycological growth.
RESULTS: Of 80 samples, 27 (33.75 per cent) had fungal growth. Sixteen patients were classified as having non-invasive fungal rhinosinusitis and 11 as having invasive fungal rhinosinusitis. The commonest clinical presentation was nasal polyposis in non-invasive fungal rhinosinusitis patients (p < 0.05) and ocular symptoms in invasive fungal rhinosinusitis patients (p < 0.05). The commonest organism was aspergillus sp. (p < 0.05) in non-invasive fungal rhinosinusitis and mucorales in invasive fungal rhinosinusitis.
CONCLUSION: There is an almost equal distribution of both invasive and non-invasive fungal rhinosinusitis, as seen in some Asian countries. Invasive fungal rhinosinusitis, while slightly uncommon when compared to non-invasive fungal rhinosinusitis, is potentially life threatening, and may require early and extensive surgical debridement. The clinical presentation of nasal polyposis was often associated with non-invasive fungal rhinosinusitis, whereas ocular symptoms were more likely to be associated with invasive fungal rhinosinusitis.
Fulltext Complete Genome Sequencing of Leptospira interrogans Isolates from Malaysia Reveals Massive Genome Rearrangement but High Conservation of Virulence-Associated Genes

Ramli SR, Bunk B, Spröer C, Geffers R, Jarek M, Bhuju S, et al.

Pathogens, 2021 Sep 15;10(9).
PMID: 34578230 DOI: 10.3390/pathogens10091198

The ability of Leptospirae to persist in environments and animal hosts but to cause clinically highly variable disease in humans has made leptospirosis the most common zoonotic disease. Considering the paucity of data on variation in complete genomes of human pathogenic Leptospirae, we have used a combination of Single Molecule Real-Time (SMRT) and Illumina sequencing to obtain complete genome sequences of six human clinical L. interrogans isolates from Malaysia. All six contained the larger (4.28-4.56 Mb) and smaller (0.34-0.395 Mb) chromosome typical of human pathogenic Leptospirae and 0-7 plasmids. Only 24% of the plasmid sequences could be matched to databases. We identified a chromosomal core genome of 3318 coding sequences and strain-specific accessory genomes of 49-179 coding sequences. These sequences enabled detailed genomic strain typing (Genome BLAST Distance Phylogeny, DNA-DNA hybridization, and multi locus sequence typing) and phylogenetic classification (whole-genome SNP genotyping). Even though there was some shared synteny and collinearity across the six genomes, there was evidence of major genome rearrangement, likely driven by horizontal gene transfer and homologous recombination. Mobile genetic elements were identified in all strains in highly varying numbers, including in the rfb locus, which defines serogroups and contributes to immune escape and pathogenesis. On the other hand, there was high conservation of virulence-associated genes including those relating to sialic acid, alginate, and lipid A biosynthesis. These findings suggest (i) that the antigenic variation, adaption to various host environments, and broad spectrum of virulence of L. interrogans are in part due to a high degree of genomic plasticity and (ii) that human pathogenic strains maintain a core set of genes required for virulence.
Fulltext High proportion of knowlesi malaria in recent malaria cases in Malaysia

Yusof R, Lau YL, Mahmud R, Fong MY, Jelip J, Ngian HU, et al.

Malar J, 2014;13:168.
PMID: 24886266 DOI: 10.1186/1475-2875-13-168

Plasmodium knowlesi is a simian parasite that has been recognized as the fifth species causing human malaria. Naturally-acquired P. knowlesi infection is widespread among human populations in Southeast Asia. The aim of this epidemiological study was to determine the incidence and distribution of malaria parasites, with a particular focus on human P. knowlesi infection in Malaysia.
Fulltext Phylogeographic Evidence for 2 Genetically Distinct Zoonotic Plasmodium knowlesi Parasites, Malaysia

Yusof R, Ahmed MA, Jelip J, Ngian HU, Mustakim S, Hussin HM, et al.

Emerg Infect Dis, 2016 Aug;22(8):1371-80.
PMID: 27433965 DOI: 10.3201/eid2208.151885

Infections of humans with the zoonotic simian malaria parasite Plasmodium knowlesi occur throughout Southeast Asia, although most cases have occurred in Malaysia, where P. knowlesi is now the dominant malaria species. This apparently skewed distribution prompted an investigation of the phylogeography of this parasite in 2 geographically separated regions of Malaysia, Peninsular Malaysia and Malaysian Borneo. We investigated samples collected from humans and macaques in these regions. Haplotype network analyses of sequences from 2 P. knowlesi genes, type A small subunit ribosomal 18S RNA and cytochrome c oxidase subunit I, showed 2 genetically distinct divergent clusters, 1 from each of the 2 regions of Malaysia. We propose that these parasites represent 2 distinct P. knowlesi types that independently became zoonotic. These types would have evolved after the sea-level rise at the end of the last ice age, which separated Malaysian Borneo from Peninsular Malaysia.