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Trend and pattern of melioidosis seropositivity among suspected patients in Malaysia 2015 - 2019

Hadi FS, Ghazali S, Ahmad N, Ramli SR

Trop Biomed, 2021 Dec 01;38(4):561-567.
PMID: 35001922 DOI: 10.47665/tb.38.4.099

Melioidosis is a disease of public health importance associated with high case-fatality rates in animals and humans caused by Burkholderia pseudomallei (B. pseudomallei ), a gram- negative bacterium that lives in tropical soil environments. This study determined the seropositivity for melioidosis among patients admitted to healthcare centres in Malaysia, from 2015 to 2019 and identified factors related to it. A total of 26,665 serum samples of suspected melioidosis patients from Malaysia hospitals were tested for IgM against B. pseudomallei. About 16.4% of total samples were seropositive and majority of them were 55 years old and above. However, younger people aged less than 15 years old were the most susceptible to the infection (AOR 3.04, p <0.001, 95% CI: 2.73, 3.39). Melioidosis infection was the highest in Sarawak (15.1%) while Perlis was the least exposed to the infection (0.9%). Further analyses showed that patients with chronic lung disease (adjusted OR: 4.03, p < 0.001, 95% CI: 2.77, 5.86) were more susceptible to melioidosis infection. In conclusion, although serology testing is not a gold standard test in diagnosing melioidosis, it has been used as a tool in treatment monitoring and disease surveillance among patients and at-risk community in the endemic hot-spots regions.
A case of mycotic aneurysm due to Burkholderia pseudomallei

Ding CH, Hussin S, Tzar MN, Rahman MM, Ramli SR

Pak J Med Sci, 2013 Apr;29(2):666-8.
PMID: 24353601

Burkholderia pseudomallei is an free-living gram-negative bacterium causing melioidosis and is endemic in Southeast Asia. A 56-year-old diabetic construction worker with a 1-month history of abdominal pain and 1-day history of high-grade fever was found to have a left non-dissecting infrarenal mycotic aortic aneurysm by abdominal computerized tomography scan. Bacteriological examination of his blood yielded Burkholderia pseudomallei. The patient was treated with right axillo-bifemoral bypass with excision of aneurysm and high-dose intravenous ceftazidime for two weeks, followed by oral trimethoprim/sulfamethoxazole and oral doxycycline for a minimum of five months.
Fulltext A Severe Case of Arthrographis kalrae Keratomycosis

Ramli SR, Francis AL, Yusof Y, Khaithir TM

Case Rep Infect Dis, 2013;2013:851875.
PMID: 24377053 DOI: 10.1155/2013/851875

A 52-year-old man with diabetes developed a unilateral central corneal ulcer after accidental foreign body inoculation. He complained of pain and loss of visual acuity in the injured eye, which displayed redness and edema and eventually discharged pus. A corneal scraping from the left eye orbit revealed fungal elements, and cultures of the material grew a fungus. The isolate was identified as Arthrographis kalrae based on gross and microscopic morphologies. The patient received amphotericin B intravenously and itraconazole orally. The wound healed following surgical intervention, but the patient lost the use of his left eye.
Fulltext Screening and detection of heterogenous vancomycin intermediate Staphylococcus aureus in Hospital Kuala Lumpur Malaysia, using the glycopeptide resistance detection Etest and population analysis profiling

Ramli SR, Neoh HM, Aziz MN, Hussin S

Infect Dis Rep, 2012 Jan 2;4(1):e20.
PMID: 24470927 DOI: 10.4081/idr.2012.e20

In a 3-month study done in Hospital Kuala Lumpur (HKL), 7 out of 320 methicillin resistant Staphylococcus aureus isolates were confirmed as heterogeneous vancomycin intermediate S. aureus (hVISA) using the glycopeptide resistance detection e-test and population analysis, giving a prevalence rate of 2.19%. This is the first report of hVISA in Malaysia.
Cryptococcus humicolus meningitis: first case report in Malaysia

Ramli SR, Leong MC, Khaithir TM, Aziz MN, Loons LC, Rafia MH

Southeast Asian J. Trop. Med. Public Health, 2012 Sep;43(5):1212-7.
PMID: 23431829

We report a case of Cryptococcus humicolus meningitis complicated by communicating hydrocephalus in an apparently immunocompetent 49-year-old psychiatric patient from a nursing home. He presented with a history of poor oral intake, weight loss, headache, vomiting, blurred vision, frequent falls and unsteady gait for the previous three months. He had a history of chronic cough, productive of whitish sputum for the previous month but no hemoptysis. Cerebrospinal fluid culture was positive for Cryptococcus humicolus. He was treated with intravenous amphotericin B and oral fluconazole and had clinical and microbiological improvement after three weeks of treatment. Unfortunately, the patient acquired nosocomial methicillin-resistant Staphylococcus aureus infection and died due to overwhelming sepsis.
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 Discovery of Leptospira spp. seroreactive peptides using ORFeome phage display

Ramli SR, Moreira GMSG, Zantow J, Goris MGA, Nguyen VK, Novoselova N, et al.

PLoS Negl Trop Dis, 2019 01;13(1):e0007131.
PMID: 30677033 DOI: 10.1371/journal.pntd.0007131

BACKGROUND: Leptospirosis is the most common zoonotic disease worldwide. The diagnostic performance of a serological test for human leptospirosis is mainly influenced by the antigen used in the test assay. An ideal serological test should cover all serovars of pathogenic leptospires with high sensitivity and specificity and use reagents that are relatively inexpensive to produce and can be used in tropical climates. Peptide-based tests fulfil at least the latter two requirements, and ORFeome phage display has been successfully used to identify immunogenic peptides from other pathogens.
METHODOLOGY/PRINCIPAL FINDINGS: Two ORFeome phage display libraries of the entire Leptospira spp. genomes from five local strains isolated in Malaysia and seven WHO reference strains were constructed. Subsequently, 18 unique Leptospira peptides were identified in a screen using a pool of sera from patients with acute leptospirosis. Five of these were validated by titration ELISA using different pools of patient or control sera. The diagnostic performance of these five peptides was then assessed against 16 individual sera from patients with acute leptospirosis and 16 healthy donors and was compared to that of two recombinant reference proteins from L. interrogans. This analysis revealed two peptides (SIR16-D1 and SIR16-H1) from the local isolates with good accuracy for the detection of acute leptospirosis (area under the ROC curve: 0.86 and 0.78, respectively; sensitivity: 0.88 and 0.94; specificity: 0.81 and 0.69), which was close to that of the reference proteins LipL32 and Loa22 (area under the ROC curve: 0.91 and 0.80; sensitivity: 0.94 and 0.81; specificity: 0.75 and 0.75).
CONCLUSIONS/SIGNIFICANCE: This analysis lends further support for using ORFeome phage display to identify pathogen-associated immunogenic peptides, and it suggests that this technique holds promise for the development of peptide-based diagnostics for leptospirosis and, possibly, of vaccines against this pathogen.