MyMedR

Fulltext Serodiagnosis of melioidosis in Malaysia

Vadivelu J, Puthucheary SD, Gendeh GS, Parasakthi N

Singapore Med J, 1995 Jun;36(3):299-302.

Current diagnosis of melioidosis is based on bacterial culture and/or serology which is becoming increasingly useful. An IgM-ELISA using heat-killed whole cells of Pseudomonas pseudomallei was developed and compared with an indirect haemagglutination technique (IHAT) and an indirect immunofluorescent technique(IFAT). The IgM-ELISA using a P:N ratio of > or = 2 had a sensitivity of 91% and a specificity of 96%. All 3 assays were further used in a seroepidemiological survey amongst different groups of patients and healthy individuals. It was found that the IFAT performed better than the IHAT, detecting antibodies to P. pseudomallei in 6% of diabetics, 5% of pyrexics, 8% of pregnant women and 3% of farmers. For the same groups the IgM-ELISA detected antibodies in 1% of pyrexics, 8% of pregnant women and a further 14% of farmers. The IgM-ELISA was found to be sensitive and useful for the serological diagnosis of acute melioidosis.

Fulltext Sequence polymorphism and PCR-restriction fragment length polymorphism analysis of the flagellin gene of Burkholderia pseudomallei

Tay ST, Cheah PC, Puthucheary SD

J Clin Microbiol, 2010 Apr;48(4):1465-7.

PMID: 20089759 DOI: 10.1128/JCM.01131-09

Four flagellin allelic types (I to IV) of Burkholderia pseudomallei were identified based on their sequence variation and restriction fragment length polymorphism (RFLP) analysis of the amplified flagellin gene. Flagellin allelic type I was the most predominantly (75.0%) found among the 100 clinical isolates of B. pseudomallei investigated in this study.

Septicaemic melioidosis: a review of 50 cases from Malaysia

Puthucheary SD, Parasakthi N, Lee MK

Trans R Soc Trop Med Hyg, 1992 11 1;86(6):683-5.

PMID: 1287945

Fifty cases of septicaemic melioidosis were reviewed. There was a preponderance of disease among males (male:female ratio 3.2:1) and those aged over 30 years. The presenting clinical features were very varied and not pathognomonic, ranging from fever, cough and septicaemia to fulminant septicaemia and shock. Pulmonary involvement was recorded in 58% of the patients. Skin and soft tissue sepsis was seen in 24%, but many had signs and symptoms of multiorgan involvement. Associated underlying illness was identified in 76% of patients, diabetes mellitus being the commonest (38%), while 34% had more than one predisposing factor. The mortality of 65% in our series is a reflection of the less than satisfactory status of the diagnosis and therapy of septicaemic melioidosis. Only 24% of our patients received appropriate empirical antibiotic therapy. A high index of suspicion of melioidosis in endemic areas and the use of appropriate empirical antimicrobial therapy would be a step towards reducing the high mortality rate.

Sensitivity of Staphylococci to antibiotics

Puthucheary SD, Chen ST, Dugdale AE

Med J Malaysia, 1972 Jun;26(4):262-265.

PMID: 35158504

No abstract available.

Salmonellosis in persons infected with human immunodeficiency virus: a report of seven cases from Malaysia

Puthucheary SD, Ng KP, Hafeez A, Raja NS, Hassan HH

Southeast Asian J. Trop. Med. Public Health, 2004 Jun;35(2):361-5.

PMID: 15691137

Persons infected with human immunodeficiency virus (HIV) have an increased risk of salmonellosis when compared to the general population. We describe seven such patients with Salmonella bacteremia, of whom two had recurrent salmonellosis. In the latter two cases the infection was unusually severe, characterized by widespread infection, bacteremia and relapse, despite standard antimicrobial therapy. HIV-infected individuals will benefit from education on the source of Salmonella, mode of acquisition and prevention through safe food handling and food preparation practices. Because of the difficulty of eradicating Salmonella infection in patients with acquired immunodeficiency syndrome, long-term suppressive treatment with antimicrobials is warranted.

Salmonella meningitis and its complications in infants

Lee WS, Puthucheary SD, Omar A

J Paediatr Child Health, 1999 Aug;35(4):379-82.

PMID: 10457297

OBJECTIVE: To review the presenting features, complications and outcome of infants with Salmonella meningitis.

METHODOLOGY: Retrospective review of all cultures of cerebrospinal fluid positive for bacteria in children below 12 years of age, processed at the Department of Medical Microbiology, University of Malaya Medical Centre, Kuala Lumpur from 1973 to 1997. Records of all cases positive for Salmonella species were retrieved and studied.

RESULTS: Thirteen infants aged 3 days to 9 months with Salmonella meningitis were included. The median age of onset of symptoms was 4 months. The clinical and laboratory features were similar to other causes of bacterial meningitis. Salmonella enteritidis was the commonest serotype isolated. Nine infants developed fits, six of which were difficult to control. Other complications noted were hydrocephalus (five), subdural effusions (four), empyema (three), ventriculitis (two), intracranial haemorrhage and cerebral abscess (one each). The use of ampicillin and/or chloramphenicol and inadequate duration of therapy resulted in recrudescence or relapse in five infants. The overall mortality was 18%. The presence of empyema, intracerebral abscess, ventriculitis, hydrocephalus, and intracranial haemorrhage were associated with adverse neurodevelopmental sequelae or death. More than half of those who survived had normal long-term outcome.

CONCLUSION: Infants who developed neurological complications as a result of Salmonella meningitis had significant mortality and adverse long-term neurodevelopment outcome.

Rotavirus and other enteropathogens in childhood acute diarrhoea: a study of two centres in Malaysia

Lee WS, Rajasekaran G, Pee S, Karunakaran R, Hassan HH, Puthucheary SD

J Paediatr Child Health, 2006 Sep;42(9):509-14.

PMID: 16925536

To study the role of rotavirus in children hospitalised for acute gastroenteritis (AGE) in two urban hospitals in Malaysia.

Ribotyping and DNA macrorestriction analysis of isolates of Burkholderia pseudomallei from cases of melioidosis in Malaysia

Vadivelu J, Puthucheary SD, Mifsud A, Drasar BS, Dance DA, Pitt TI

Trans R Soc Trop Med Hyg, 1997 5 1;91(3):358-60.

PMID: 9231217

Forty-nine isolates of Burkholderia pseudomallei from sporadic cases of melioidosis in Malaysia over the past 18 years were examined by BamHI ribotyping and pulsed-field gel electrophoresis (PFGE) of XbaI digests of total deoxyribonucleic acid (DNA). Twenty-four patients had septicaemic melioidosis with a mortality of 70%; mortality in the non-septicaemic disease was 16%. Five ribotype patterns were identified, 2 of which accounted for 90% of all isolates. PFGE revealed a number of different strains within these ribotypes, but some pairs of isolates from unrelated cases gave closely similar DNA profiles. These results are in agreement with Australian studies which showed a high prevalence of a few ribotypes of B. pseudomallei which are further divisible by genotyping, in areas where melioidosis is endemic.

Rhodococcus equi--an emerging human pathogen in immunocompromized hosts: a report of four cases from Malaysia

Puthucheary SD, Sangkar V, Hafeez A, Karunakaran R, Raja NS, Hassan HH

Southeast Asian J. Trop. Med. Public Health, 2006 Jan;37(1):157-61.

PMID: 16771229

Rhodococcus equi, a recognized pathogen in horses, is emerging as a human opportunistic pathogen, especially in immunocompromized hosts. We describe four immunocompromized patients who had serious R. equi infections with an overall mortality of 75%. The natural habitat of R. equi is soil, particularly soil contaminated with animal manure. Necrotizing pneumonia is the commonest form of infection but extrapulmonary infections, such as wound infections and subcutaneous abscess, have also been described in humans. R. equi is cultured easily in ordinary non-selective media. Large, smooth, irregular colonies appear within 48 hours. It is a facultative, intracellular, nonmotile, non-spore forming, gram-positive coccobacillus, which is weakly acid-fast staining and bears a similarity to diphtheroids. It forms a salmon-colored pigment usually after 48 hours incubation. A particular characteristic of this organism is that it undergoes synergistic hemolysis with some bacteria on sheep blood agar. R. equi may be misidentified as diphtheroids, Mycobacterium species, or Nocardia. In vitro R. equi is usually susceptible to erythromycin, ciprofloxacin, vancomycin, aminoglycosides, rifampin, imipenem and meropenem. The organism can be difficult to eradicate, making treatment challenging. Increased awareness of the infection may help with early diagnosis and timely treatment.

Fulltext Retrospective study of Aeromonas infection in a Malaysian urban area: a 10-year experience

Lee WS, Puthucheary SD

Singapore Med J, 2001 Feb;42(2):057-60.

PMID: 11358191

To describe the patterns of isolation of Aeromonas spp. and the resulting spectrum of infection, intestinal and extra-intestinal,from infants and children in an urban area in a hot and humid country from SoutheastAsia.

Fulltext Reduced susceptibility of Malaysian clinical isolates of Burkholderia pseudomallei to ciprofloxacin

Liew FY, Tay ST, Puthucheary SD

Trop Biomed, 2011 Dec;28(3):646-50.

PMID: 22433895 MyJurnal

Ciprofloxacin, a quinolone with good intracellular penetration may possibly be used for treatment of melioidosis caused by Burkholderia pseudomallei, but problems with resistance may be encountered. Amino acid substitutions in gyrA/gyrB have given rise to fluoroquinolone resistance in various microorganisms. Using published primers for gyrA and gyrB, PCR was performed on 11 isolates of B. pseudomallei with varying degrees of sensitivity to ciprofloxacin, followed by DNA sequencing to detect possible mutations. Results showed an absence of any point mutation in either gene. Local isolates have yet to develop full resistance to ciprofloxacin and probably other mechanisms of resistance may have been involved in the decreased sensitivity to ciprofloxacin.

Fulltext Rapid identification of melioidosis agent by an insulated isothermal PCR on a field-deployable device

Chua KH, Tan EW, Chai HC, Puthucheary SD, Lee PC, Puah SM

PeerJ, 2020;8:e9238.

PMID: 32518734 DOI: 10.7717/peerj.9238

Background: Burkholderia pseudomallei causes melioidosis, a serious illness that can be fatal if untreated or misdiagnosed. Culture from clinical specimens remains the gold standard but has low diagnostic sensitivity.

Method: In this study, we developed a rapid, sensitive and specific insulated isothermal Polymerase Chain Reaction (iiPCR) targeting bimA gene (Burkholderia Intracellular Motility A; BPSS1492) for the identification of B. pseudomallei. A pair of novel primers: BimA(F) and BimA(R) together with a probe were designed and 121 clinical B. pseudomallei strains obtained from numerous clinical sources and 10 ATCC non-targeted strains were tested with iiPCR and qPCR in parallel.

Results: All 121 B. pseudomallei isolates were positive for qPCR while 118 isolates were positive for iiPCR, demonstrating satisfactory agreement (97.71%; 95% CI [93.45-99.53%]; k = 0.87). Sensitivity of the bimA iiPCR/POCKIT assay was 97.52% with the lower detection limit of 14 ng/µL of B. pseudomallei DNA. The developed iiPCR assay did not cross-react with 10 types of non-targeted strains, indicating good specificity.

Conclusion: This bimA iiPCR/POCKIT assay will undoubtedly complement other methodologies used in the clinical laboratory for the rapid identification of this pathogen.

Quorum sensing in Aeromonas species isolated from patients in Malaysia

Chan KG, Puthucheary SD, Chan XY, Yin WF, Wong CS, Too WS, et al.

Curr Microbiol, 2011 Jan;62(1):167-72.

PMID: 20544198 DOI: 10.1007/s00284-010-9689-z

Bacterial quorum sensing signal molecules called N-acylhomoserine lactone (AHL) controls the expression of virulence determinants in many Gram-negative bacteria. We determined AHL production in 22 Aeromonas strains isolated from various infected sites from patients (bile, blood, peritoneal fluid, pus, stool and urine). All isolates produced the two principal AHLs, N-butanoylhomoserine lactone (C4-HSL) and N-hexanoyl homoserine lactone (C6-HSL). Ten isolates also produced additional AHLs. This report is the first documentation of Aeromonas sobria producing C6-HSL and two additional AHLs with N-acyl side chain longer than C(6). Our data provides a better understanding of the mechanism(s) of this environmental bacterium emerging as a human pathogen.

Purulent meningitis in childhood

Lee EI, Khoo BH, Puthucheary SD, Thong ML

Med J Malaysia, 1977 Dec;32(2):114-9.

PMID: 26858

Pulsed-field gel electrophoresis of multidrug-resistant and -sensitive strains of Pseudomonas aeruginosa from a Malaysian hospital

Thong KL, Lai KS, Ganeswrie R, Puthucheary SD

Jpn J Infect Dis, 2004 Oct;57(5):206-9.

PMID: 15507777

Over a period of 6 months from January to June 2002, an unusual increase in the isolation of highly resistant Pseudomonas aeruginosa strains was observed in the various wards and intensive care units of a large general hospital in Johor Bahru, Malaysia. An equal number of multidrug resistant (MDR) and drug-susceptible strains were collected randomly from swabs, respiratory specimens, urine, blood, cerebral spinal fluid, and central venous catheters to determine the clonality and genetic variation of the strains. Macrorestriction analysis by pulsed-field gel electrophoresis showed that the 19 MDR strains were genetically very homogenous; the majority showed the dominant profile S1 (n = 10), the rest very closely related profiles S1a (n = 1), S2 (n = 4), and S2a (n = 3), indicating the endemicity of these strains. In contrast, the 19 drug-sensitive strains isolated during the same time period were genetically more diverse, showing 17 pulsed-field profiles (F = 0.50-1.00), and probably derived from the patients themselves. The presence of the MDR clone poses serious therapeutic problems as it may become endemic in the hospital and give rise to future clonal outbreaks. There is also the potential for wider geographical spread.

Proteomic profiling of clinical and environmental strains of Pseudomonas aeruginosa

Liew SM, Puthucheary SD, Rajasekaram G, Chai HC, Chua KH

Mol Biol Rep, 2021 Mar;48(3):2325-2333.

PMID: 33728559 DOI: 10.1007/s11033-021-06262-8

Pseudomonas aeruginosa is a ubiquitous bacterium, which is able to change its physiological characteristics in response to different habitats. Environmental strains are presumably less pathogenic than clinical strains and whether or not the clinical strains originate from the environment or through inter-host transmission remains poorly understood. To minimize the risk of infection, a better understanding of proteomic profiling of P. aeruginosa is necessary for elucidating the correlation between environmental and clinical strains. Based on antimicrobial susceptibility and patterns of virulence, we selected 12 clinical and environmental strains: (i) environmental, (ii) multidrug resistant (MDR) clinical and (iii) susceptible clinical strains. Whole-cell protein was extracted from each strain and subjected to two-dimensional differential gel electrophoresis (2-D DIGE) and liquid chromatography tandem mass spectrometry quadrupole time-of-flight (LC-MS QTOF). All 12 strains were clustered into 3 distinct groups based on their variance in protein expression. A total of 526 matched spots were detected and four differentially expressed protein spots (p < 0.05) were identified and all differential spots were downregulated in MDR strain J3. Upregulation of chitin binding and BON domain proteins was present in the environmental and some MDR strains, whereas the clinical strains exhibited distinct proteomic profiles with increased expression of serine protein kinase and arginine/ornithine transport ATP-binding proteins. Significant difference in expression was observed between susceptible clinical and MDR strains, as well as susceptible clinical and environmental strains. Transition from an environmental saprophyte to a clinical strain could alter its physiological characteristics to further increase its adaptation.

Fulltext Potential immunogenic polypeptides of Burkholderia pseudomallei identified by shotgun expression library and evaluation of their efficacy for serodiagnosis of melioidosis

Puah SM, Puthucheary SD, Chua KH

Int J Med Sci, 2013;10(5):539-47.

PMID: 23532805 DOI: 10.7150/ijms.5516

The search for novel immunogenic polypeptides to improve the accuracy and reliability of serologic diagnostic methods for Burkholderia pseudomallei infection is ongoing. We employed a rapid and efficient approach to identify such polypeptides with sera from melioidosis patients using a small insert genomic expression library created from clinically confirmed local virulent isolates of B. pseudomallei. After 2 rounds of immunoscreening, 6 sero-positive clones expressing immunogenic peptides were sequenced and their identities were: benzoate 1,2-dioxygenase beta subunit, a putative 200 kDa antigen p200, phosphotransferase enzyme family protein, short chain dehydrogenase and 2 hypothetical proteins. These immunogens were then transferred to an ELISA platform for further large scale screening. By combining shotgun expression library and ELISA assays, we identified 2 polypeptides BPSS1904 (benzoate 1,2-dioxygenase beta subunit) and BPSL3130 (hypothetical protein), which had sensitivities of 78.9% and 79.4% and specificities of 88.1% and 94.8%, respectively in ELISA test, thus suggesting that both are potential candidate antigens for the serodiagnosis of infections caused by B. pseudomallei.

Possible virulence factors involved in bacteraemia caused by Aeromonas hydrophila

Vadivelu J, Puthucheary SD, Phipps M, Chee YW

J Med Microbiol, 1995 Mar;42(3):171-4.

PMID: 7884797

Eighteen strains of Aeromonas hydrophila from patients with bacteraemia were investigated for possible virulence factors. Cytotoxin and haemolysin were produced by all strains, whereas cholera toxin-like factor was produced by 33% of strains only. Enterotoxin production was not detected. Haemagglutination of guinea-pig, fowl and rabbit erythrocytes was demonstrated by 83%, 67% and 61% of strains, respectively. Fucose- and mannose-sensitive haemagglutinins were predominant. None of the strains agglutinated sheep erythrocytes. Extrachromosomal DNA was detected in 17 strains, 16 of which had a plasmid (3.6-5.1 MDa), the majority being between 4.6 and 5.1 MDa.

Fulltext Phenotypic and Genetic Diversity of Aeromonas Species Isolated from Fresh Water Lakes in Malaysia

Khor WC, Puah SM, Tan JA, Puthucheary SD, Chua KH

PLoS One, 2015;10(12):e0145933.

PMID: 26710336 DOI: 10.1371/journal.pone.0145933

Gram-negative bacilli of the genus Aeromonas are primarily inhabitants of the aquatic environment. Humans acquire this organism from a wide range of food and water sources as well as during aquatic recreational activities. In the present study, the diversity and distribution of Aeromonas species from freshwater lakes in Malaysia was investigated using glycerophospholipid-cholesterol acyltransferase (GCAT) and RNA polymerase sigma-factor (rpoD) genes for speciation. A total of 122 possible Aeromonas strains were isolated and confirmed to genus level using the API20E system. The clonality of the isolates was investigated using ERIC-PCR and 20 duplicate isolates were excluded from the study. The specific GCAT-PCR identified all isolates as belonging to the genus Aeromonas, in agreement with the biochemical identification. A phylogenetic tree was constructed using the rpoD gene sequence and all 102 isolates were identified as: A. veronii 43%, A. jandaei 37%, A. hydrophila 6%, A. caviae 4%, A. salmonicida 2%, A. media 2%, A. allosaccharophila 1%, A. dhakensis 1% and Aeromonas spp. 4%. Twelve virulence genes were present in the following proportions--exu 96%, ser 93%, aer 87%, fla 83%, enolase 70%, ela 62%, act 54%, aexT 33%, lip 16%, dam 16%, alt 8% and ast 4%, and at least 2 of these genes were present in all 102 strains. The ascV, aexU and hlyA genes were not detected among the isolates. A. hydrophila was the main species containing virulence genes alt and ast either present alone or in combination. It is possible that different mechanisms may be used by each genospecies to demonstrate virulence. In summary, with the use of GCAT and rpoD genes, unambiguous identification of Aeromonas species is possible and provides valuable data on the phylogenetic diversity of the organism.

Nosocomial outbreak of Enterobacter gergoviae bacteraemia in a neonatal intensive care unit

Ganeswire R, Thong KL, Puthucheary SD

J Hosp Infect, 2003 Apr;53(4):292-6.

PMID: 12660126

A nosocomial outbreak of bacteraemia, caused by Enterobacter gergoviae infected 11 babies, nine of whom were premature, and was investigated in the neonatal intensive care unit (NICU) of a general hospital in Johor Bahru, Malaysia. The strain that was isolated from the babies was also isolated from the dextrose saline used for the dilution of parenteral antibiotics and from the hands of a healthcare worker on duty in the nursery. Pulsed-field gel electrophoresis (PFGE) of Xba I-digested chromosomal DNA confirmed a possible cross-contamination of parenteral dextrose saline and the healthcare worker. Prompt and effective control measures were initiated within NICU and the nosocomial infection of E. gergoviae was brought to an abrupt end. To the best of our knowledge, this is the first documented outbreak of E. gergoviae in the NICU in a hospital in the state of Johor, Malaysia.

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