Objective: To characterise a collection of pili-carrying and none pili-carrying pneumococcal isolates of clinical origin for serotypes, antibiotic resistance and genotype. Methods: In total, 42 clinical isolates were collected between October 2017 and December 2019. Those isolates were analysed for antimicrobial susceptibility, serotype distribution, detection of pneumococcal virulence and pilus genes. Multilocus sequence typing was performed only for piliated isolates, followed by phylogenetic analysis. Results: The common isolation sites among the pneumococcal isolates were tracheal aspirate (28.6%), blood (26.2%), and sputum (23.8%). Fifty percent isolates were resistant to erythromycin, tetracycline (50.0%) and trimethoprim-sulfamethoxazole (43.0%). The most frequent were serotypes 19F (28.6%), 6A/B (23.8%) and 19A (14.3%). Piliated isolates were detected in a small proportion (33.3%); 64.3% were multidrug-resistant. ST320 was the prevalent sequence type among the piliated isolates and genetically related to the Pneumococcal Molecular Epidemiology Network clones Taiwan 19F -14 (CC271). In the phylogenetic analysis, some piliated isolates showed a close association having similar ST320, carrying serotype 19A and both pilus genes indicating their clonal spread. Conclusions: Pneumococcal lineages of piliated isolates have been globally disseminated and pili could have played a role in the spread of antibiotic resistant clones.

Introduction: Limited studies have been documented on the presence of pathogenic Leptospira in public markets serving the community in sub-districts of Selangor. The aim of this study was to detect the presence of pathogenic Leptospira in rats using a gene encoding an outer membrane lipoprotein LipL32. Methods: Polymerase chain reaction (PCR) was performed using LipL32 primers on sixty kidney samples of rats trapped at two locations of study; Pasar Borong Selangor in Seri Kembangan and Pasar Basah Bandar Baru Bangi in Bangi. Results: Out of 60 samples analysed, 36.7% were positive for the presence of LipL32. All positive samples highly matched (>94%) nucleotide sequence for LipL32 of pathogenic Leptospira and related to the pathogens through phylogenetic analysis. Conclusion: The detection of LipL32 indicates the potential presence of pathogenic Leptospira species at public markets. Although only 60 rats were successfully trapped, the rats are mobile and might further transmit the pathogenic organisms to other areas.

Background: Streptococcus pyogenes has a variety of virulence factors and the predominant invasive strains differ according to specific emm types and geographical orientation. Although emm typing is commonly used as the gold standard method for the molecular characterisation, multilocus sequence typing (MLST) has become an important tool for comparing the genetic profiles globally. This study aimed to screen selected virulence genes from invasive and non-invasive clinical samples and to characterise the molecular epidemiology by emm typing and MLST methods. Materials and Methods: A total of 42 S. pyogenes isolates from invasive and non-invasive samples collected from two different tertiary hospitals were investigated for the distribution of virulence factors and their molecular epidemiology by emm and multilocus sequence typing methods. Detection of five virulence genes (speA, speB, speJ, ssa and sdaB) was performed using multiplex polymerase chain reaction (PCR) using the standard primers and established protocol. Phylogenetic tree branches were constructed from sequence analysis utilised by neighbour joining method generated from seven housekeeping genes using MEGA X software. Results: Multiplex PCR analysis revealed that sdaB/speF (78.6%) and speB (61.9%) were the predominant virulence genes. Regardless of the type of invasiveness, diverse distribution of emm types/subtypes was noted which comprised of 27 different emm types/subtypes. The predominant emm types/subtypes were emm63 and emm18 with each gene accounted for 11.8% whereas 12% for each gene was noted for emm28, emm97.4 and emm91. The MLST revealed that the main sequence type (ST) in invasive samples was ST402 (17.7%) while ST473 and ST318 (12% for each ST) were the major types in non-invasive samples. Out of 18 virulotypes, Virulotype A (five genes, 55.6%) and Virulotype B (two genes, 27.8%) were the major virulotypes found in this study. Phylogenetic analysis indicated the presence of seven different clusters of S. pyogenes. Interestingly, Cluster VI showed that selected emm/ST types such as emm71/ST318 (n=2), emm70.1/ST318 (n=1), emm44/ST31 (n=1) and emm18/ST442 (n=1) have clustered within a common group (Virulotype A) for both hospitals studied. Conclusion: The present study showed that group A streptococcci (GAS) are genetically diverse and possess virulence genes regardless of their invasiveness. Majority of the GAS exhibited no restricted pattern of virulotypes except for a few distinct clusters. Therefore, it can be concluded that virulotyping is partially useful for characterising a heterogeneous population of GAS in hospitals.

Introduction: Coagulase-negative staphylococci (CoNS) is often considered as a culture contaminants but it can potentially be pathogenic to patients with risk factors. A combination of species identification and clinical criteria has been suggested in determining true CoNS bacteraemia. Objectives: To identify the species distribution, antibiotic susceptibility patterns and clinical profiles of CoNS isolated from blood cultures among paediatric patients in Hospital Kuala Lumpur (HKL). Methods: This study involved CoNS isolation from blood cultures of paediatric in-patients of the Paediatric Institute HKL. Isolates were identified to species level using Analytical Profile Index Staph identification strips and antimicrobial susceptibility pattern following Kirby-Bauer Disc Diffusion method. The clinical profiles of patients were obtained from their medical records. Results: Eleven CoNS species were identified from 148 isolates. Staphylococcus epidermidis was the most frequent species isolated (67.6%). The majority of the isolates showed resistance to penicillin (85.8%); while 70.3% were methicillin-resistant (MR) CoNS, which demonstrated a significant association with true infection (p=0.021). Predictors for significant CoNS infection included thrombocytopaenia, presence of predisposing factors, nosocomial infection, blood collected from peripheral vein, and CoNS isolated from two consecutive blood cultures. The most common predisposing factors for the isolation of CoNS were the presence of peripheral (54.1%) and central venous catheters (35.1%). Conclusion: CoNS can cause significant bloodstream infections. The isolation of CoNS from blood cultures should be carefully interpreted by considering the predictive factors. Local data regarding predictive factors of patients with culture-positive CoNS, species distribution and antimicrobial susceptibility pattern are useful to determine the significance of blood culture results and care management of patients

Suppurative BCG lymphadenitis can easily be overlooked, as it mimics other diseases such as tuberculous lymphadenitis. A case of a three-month old female infant who received the BCG vaccination at birth presented with isolated left axillary mass at two months of age. She was initially treated as lymph node abscess but was referred to the hospital due to the increasing size of the swelling. Needle aspiration was done and the microbiology analysis came out positive for acid-fast bacilli. She was planned for syrup isoniazid; however, the management team withheld treatment until they were certain of the identity of the bacteria. The bacteria was confirmed by the molecular method to be Mycobacterium bovis BCG strain. The case report highlights the importance of the microbiology investigations for appropriate management in this case.

Despite pulmonary cryptococcosis showing good response to antifungal treatment, making accurate diagnosis in a timely manner remains a challenge. Tissue for fungal culture is less sensitive, nonspecific, and time-consuming to allow for prompt diagnosis. We herein report a case of 31-year-old woman with no known medical illness but presented with history of cough and haemoptysis for two months. The chest X-ray and bronchoscopy revealed left lung mass. While tuberculosis showed negative result and no growth observed in the tissue culture, the histopathological examination (HPE) finding was suggestive of fungal infection. The formalin-fixed paraffin-embedded (FFPE) tissue was sent for molecular testing, which revealed Cryptococcus neoformans. This report emphasises on the advantages and limitations of polymerase chain reaction (PCR) as an alternative method to confirm the diagnosis in cases of culture-negative fungal infection.

Automated microbial detection system (AMDs) are design to detect early growth of bacterial and fungal. We herein report a rare case of false positive blood culture by AMDs in Plasmodium falciparum infection. A 41-year-old previously healthy lady, with recent history of travelling to Lagos, Nigeria had presented to the casualty with history of fever and lethargy for three days. There was no malaria prophylaxis taken prior to the travelling history. Peripheral blood smear confirmed the presence of young trophozoite of Plasmodium falciparum with parasitemia of 7%. Concurrent blood culture sent was positive, however all subcultures were negative for any growth. She was treated with intravenous artesunate however succumbed to death on the day of admission due to severe falciparum infection complicated with multiorgan failure and shock. The aim of this report is to highlight, the circumstances that can trigger the false positive AMDs detection and the possible underlying mechanism.

Rhinocerebral mucormycosis is a potentially fatal and progressive angioinvasive fungal infection. It is classically described in patients with uncontrolled diabetes mellitus and hematological malignancies. This report describes a case of progressive rhinocerebral mucormycosis in a patient with poorly controlled diabetes who was on prolonged prednisolone therapy for autoimmune kidney disease. The patient, who was a female, presented to hospital with headache, orbital pain and nasal bridge swelling. Black eschar on nasal mucosae was present on admission. Later, she was started on intravenous fluconazole for the diagnosis of fungal sinusitis. Subsequently, she developed intracerebral haemorrhage complicated with transtentorial herniation. Diagnosis of rhinocerebral mucormycosis was later observed by a laboratory finding and the treatment was changed to intravenous amphotericin B. However, the patient succumbed to her illness on the 6th day of hospitalisation. This report discusses the risk factors associated with rhinocerebral mucormycosis as well as the underlying pathogenesis. This report will also highlight the importance of early diagnosis and appropriate treatment for mucormycosis to improve prognosis in patients.

We present a unique case of pneumococcal meningitis caused by serotype 15C, a non-vaccine serotype, which is long thought to be non-virulent. Our patient is a young lady with a known case of plaque psoriasis, presented with sudden onset of left oculomotor nerve palsy and severe headache two days prior to admission. The clinical features were initially mimicking of brain tumour and brain aneurysm. The diagnosis of pneumococcal meningitis was confirmed by the bacterial antigen test and genome detection using multiplex PCR from the CSF. The blood culture also grew Streptococcus pneumoniae. Serotyping was performed on the isolate using Neufeld's Quellung method and it was identified as serotype 15C. Psoriatic skin disease was identified as a potential source of this invasive infection.