MyMedR

Acute septicaemic melioidosis: a report of seven cases

Puthucheary SD, Lin HP, Yap PK

Trop Geogr Med, 1981 Mar;33(1):19-22.

A report is presented of seven patients with acute septicaemic melioidosis seen at the University Hospital, Kuala Lumpur, Malaysia, during 1976-1979. All had associated disorders which rendered them more susceptible to infection. As prognosis depends on early diagnosis it is important that this disease be considered in the differential diagnosis of a septicaemic illness in such patients from endemic areas. The treatment of choice is a combination of tetracyclines and chloramphenicol, initially used in massive doses, and continued for at least six month to prevent relapses.

Fulltext Bacteraemic enteritis due to Campylobacter jejuni

Puthucheary SD, Lin HP

Med J Malaysia, 1982 Dec;37(4):378-80.

PMID: 7167093

Campylobacter Jejuni is being increasingly recognised as a cause of bacteraemia enteritis and two infants with this condition are described. Awareness of the organism. as a possible cause of septicaemia is important because it has special growth requirements and delay in the diagnosis can be detrimental in a disease which usually only responds to erythromycin, gentamicin and chloramphenicol.

Ceftazidime and cefotaxime--the clinician's choice

Puthucheary SD, Goldsworthy PJ

Clin Ther, 1989 Mar-Apr;11(2):186-204.

PMID: 2660995

A review of two third-generation cephalosporins, ceftazidime and cefotaxime, is presented. Ceftazidime, often used as a single agent, has shown greater activity than cefotaxime against Pseudomonas aeruginosa and other Pseudomonas species, Enterobacteriaceae, Acinetobacter sp, and Enterobacter sp. It has been effective as monotherapy in the treatment of peritonitis, gynecologic infections, chronic bronchitis, and infections in patients with leukemia and granulocytopenia, as has cefotaxime when in combination with an aminoglycoside. Cefotaxime has shown good activity against most aerobic gram-negative bacilli and against Staphylococcus. It has been used in respiratory infections, urinary tract infections, and septicemia. In contrast to first-generation and most second-generation cephalosporins, third-generation cephalosporins have proven useful in some types of meningitis. Ceftazidime and cefotaxime successfully penetrate into the cerebrospinal fluid and cures of bacterial meningitis have been reported with both drugs. Both ceftazidime and cefotaxime have been successfully used in children, infants, and neonates, as well as adults. Safety profiles of ceftazidime compare favorably with those of other third-generation cephalosporins.

Fulltext Susceptibility of 57 strains Ps. pseudomallei to some new B-lactams and other antibiotics

Puthucheary SD, Parasakthi N

Med J Malaysia, 1987 Dec;42(4):248-51.

PMID: 3454397

Fifty seven strains of Pseudomonas pseudomallei were tested for in vitro susceptibility to 15 antimicrobial agents. Amongst the generally recommended antibiotics for therapy of melioidosis, only 86%, 84% and 58% of the strains were found to be sensitive to trimethoprim-sulphamethoxazole, chloramphenicol and tetracycline respectively. Of the newer B-Iactams, in descending order of activity were, ceftazidime, ceftriaxone, cefotaxime, cefoperazone and cefuroxime. But on a weight for weight basis, ceftazidime was the most active agent and as such, may be considered in the therapy of acute septicaemic melioidosis."

Fulltext A curriculum in medical ethics and medical humanities

Puthucheary SD

Med J Malaysia, 1980 Sep;35(1):86-95.

PMID: 7254006

The code of ethics derived from the Hippocratic Oath needs to be supplemented by a formal curriculum in Medical Ethics and Medical Humanities in our Medical schools. The need and justification for it, a review of the medical ethics curricula in American. European. British and Australian Universities, together with an outline of the proposed curriculum is described.

The sensitivity of staphylococci to antibiotics

Puthucheary SD, Chen ST, Dugdale AE

Med J Malaya, 1972 Jun;26(4):262-5.

PMID: 5069415

Vibrio parahaemolyticus gastroenteritis in Malaysia

Puthucheary SD

Med J Malaysia, 1973 Sep;28(1):44-6.

PMID: 4273784

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.

Fulltext Acute respiratory failure in melioidosis

Puthucheary SD, Vadivelu J, Wong KT, Ong GS

Singapore Med J, 2001 Mar;42(3):117-21.

PMID: 11405563

In melioidosis caused by Burkholderia pseudomallei, although every organ in the body may be involved, the highest mortality of 73% occurs when the respiratory system is affected. These patients invariably die of acute respiratory failure. Most of them also have underlying predisposing factors like diabetes mellitus.

Fulltext Comparison of serum F2 isoprostane levels in diabetic patients and diabetic patients infected with Burkholderia pseudomallei

Puthucheary SD, Nathan SA

Singapore Med J, 2008 Feb;49(2):117-20.

PMID: 18301838

Oxidative stress can occur in sepsis and infection, when overproduction of free radicals is not countered by the host antioxidant system, leading to impairment of host cellular functions. Various disease states are accompanied by the accumulation of 15-F2t-IsoP in biological fluids. These isoprostanes are considered as markers of oxidative stress, and inflammation and inflammatory mediators.

Fulltext Comparison by electron microscopy of intracellular events and survival of Burkholderia pseudomallei in monocytes from normal subjects and patients with melioidosis

Puthucheary SD, Nathan SA

Singapore Med J, 2006 Aug;47(8):697-703.

PMID: 16865211

Burkholderia pseudomallei (B. pseudomallei) has been shown to persist intracellularly in patients with melioidosis, until reactivated by decreasing immunocompetence. We have previously demonstrated by transmission electron microscopy, the internalisation of B. pseudomallei by human macrophages and the occurrence of phagosome-lysosome fusion.

Fulltext Campylobacter enteritis in children: clinical and laboratory findings in 137 cases

Puthucheary SD, Parasakthi N, Liew ST, Chee YW

Singapore Med J, 1994 Oct;35(5):453-6.

PMID: 7701360

One hundred and thirty-seven children with Campylobacter diarrhoea were reviewed. The predominant species was C. jejuni. Ninety-five percent of the children were below 5 years of age with 61% of these being 2-12 months old. A slight male preponderance was noted. About half the cases presented with fever and bloody diarrhoea; vomiting was seen in 28% and abdominal colic in only 8%. Moderate to severe diarrhoea was present in 48% of the children. Thirty-seven percent had a history of recent or concurrent illness. Other bacterial enteropathogens together with Campylobacter were isolated in 15% of the children. Erythromycin, the most useful drug, when indicated for Campylobacter infections, had an MIC90 of 2 mg/l with 96.2% of the strains being sensitive.

Fulltext Methicillin resistant Staphylococcus aureus--first case of bacteremia in the University Hospital, Kuala Lumpur

Puthucheary SD, Lim CT, Parasakthi N, Tan A, Lam KL

Singapore Med J, 1987 Oct;28(5):456-8.

PMID: 3433116

Fulltext Molecular characterization of putative virulence determinants in Burkholderia pseudomallei

Puah SM, Puthucheary SD, Wang JT, Pan YJ, Chua KH

ScientificWorldJournal, 2014;2014:590803.

PMID: 25215325 DOI: 10.1155/2014/590803

The Gram-negative saprophyte Burkholderia pseudomallei is the causative agent of melioidosis, an infectious disease which is endemic in Southeast Asia and northern Australia. This bacterium possesses many virulence factors which are thought to contribute to its survival and pathogenicity. Using a virulent clinical isolate of B. pseudomallei and an attenuated strain of the same B. pseudomallei isolate, 6 genes BPSL2033, BP1026B_I2784, BP1026B_I2780, BURPS1106A_A0094, BURPS1106A_1131, and BURPS1710A_1419 were identified earlier by PCR-based subtractive hybridization. These genes were extensively characterized at the molecular level, together with an additional gene BPSL3147 that had been identified by other investigators. Through a reverse genetic approach, single-gene knockout mutants were successfully constructed by using site-specific insertion mutagenesis and were confirmed by PCR. BPSL2033::Km and BURPS1710A_1419::Km mutants showed reduced rates of survival inside macrophage RAW 264.7 cells and also low levels of virulence in the nematode infection model. BPSL2033::Km demonstrated weak statistical significance (P = 0.049) at 8 hours after infection in macrophage infection study but this was not seen in BURPS1710A_1419::Km. Nevertheless, complemented strains of both genes were able to partially restore the gene defects in both in vitro and in vivo studies, thus suggesting that they individually play a minor role in the virulence of B. pseudomallei.

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.

Fulltext First Report of Extended-Spectrum β-Lactamases TEM-116 and OXA-10 in Clinical Isolates of Alcaligenes Species from Kuala Lumpur, Malaysia

Puah SM, Puthucheary SD, Chua KH

Jpn J Infect Dis, 2019 Jul 24;72(4):266-269.

PMID: 30918144 DOI: 10.7883/yoken.JJID.2018.031

There is an alarming increase in the prevalence of extended-spectrum β-lactamases (ESBLs) present mainly in Enterobacteriaceae and other nonfermenting gram-negative bacteria, such as Alcaligenes faecalis, which is the only species in that genus that is clinically relevant. We investigated Alcaligenes species from 7 cases (6 inpatients and one outpatient) at our tertiary-care hospital. Four patients had urinary tract infections, and one each had systemic lupus erythematosus, pulmonary stenosis, and diabetic ulcer. All 7 isolates were identified as Alcaligenes spp. based on their 16S rRNA gene sequences, and antibiotic susceptibility was determined using a Vitek 2 system with AST-GN87 cards. All the strains were resistant to cefazolin; 6 were resistant to trimethoprim/sulfamethoxazole; 5 manifested resistance to ampicillin/sulbactam, cefepime, tobramycin, ciprofloxacin, and nitrofurantoin; whereas 5 had multidrug resistance profiles. All the strains (7/7) expressed ESBL activity; PCR screening and sequencing showed evidence of genes blaTEM-116 (7/7) and blaOXA-10 (4/7), and we believe that this is the first report on the presence of TEM-116 and OXA-10 in an Alcaligenes spp. A combination of the 2 genes was present in 4 strains. All 7 strains were found to harbor at least one ESBL gene probably contributing to the drug resistance.

Aeromonas aquariorum clinical isolates: antimicrobial profiles, plasmids and genetic determinants

Puah SM, Puthucheary SD, Liew FY, Chua KH

Int J Antimicrob Agents, 2013 Mar;41(3):281-4.

PMID: 23312608 DOI: 10.1016/j.ijantimicag.2012.11.012

The objective of this study was to investigate the antimicrobial resistance patterns of 47 clinical isolates of Aeromonas aquariorum and to identify the presence of plasmids and the relevant antibiotic resistance genes (ARGs). Antibiotic susceptibilities were determined by the standard disc diffusion method. The presence of plasmids and ARGs was detected by gel electrophoresis and monoplex PCR. Resistance to amoxicillin/clavulanic acid (98%), amoxicillin (91%), gentamicin (13%), trimethoprim/sulfamethoxazole (11%) and kanamycin (6%) was observed, whilst no ciprofloxacin- or amikacin-resistant strains were detected. All isolates harboured plasmids with sizes ranging from ca. 2 kb to 10 kb. PCR revealed that A. aquariorum carried three β-lactam resistance genes (bla(TEM), bla(MOX) and bla(PSE)) and two sulphonamide resistance genes (sul1 and sul2). This study provides further understanding of the phenotypic and genotypic characteristics of multiresistant A. aquariorum clinical isolates.

Development of a species-specific PCR-RFLP targeting rpoD gene fragment for discrimination of Aeromonas species

Puah SM, Khor WC, Kee BP, Tan JAMA, Puthucheary SD, Chua KH

J Med Microbiol, 2018 Sep;67(9):1271-1278.

PMID: 30024365 DOI: 10.1099/jmm.0.000796

PURPOSE: The taxonomy of Aeromonas keeps expanding and their identification remains problematic due to their phenotypic and genotypic heterogeneity. In this study, we aimed to develop a rapid and reliable polymerase chain reaction-restriction fragment length polymorphism assay targeting the rpoD gene to enable the differentiation of aeromonads into 27 distinct species using microfluidic capillary electrophoresis.

METHODOLOGY: A pair of degenerate primers (Aero F: 5'-YGARATCGAYATCGCCAARCGB-3' and Aero R: 5'-GRCCDATGCTCATRCGRCGGTT-3') was designed that amplified the rpoD gene of 27 Aeromonas species. Subsequently, in silico analysis enabled the differentiation of 25 species using the single restriction endonuclease AluI, while 2 species, A. sanarelli and A. taiwanensis, required an additional restriction endonuclease, HpyCH4IV. Twelve type strains (A. hydrophila ATCC7966T, A. caviae ATCC15468T, A. veronii ATCC9071T, A. media DSM4881T, A. allosaccharophila DSM11576T, A. dhakensis DSM17689T, A. enteropelogens DSM7312T, A. jandaei DSM7311T, A. rivuli DSM22539T, A. salmonicida ATCC33658T, A. taiwanensis DSM24096T and A. sanarelli DSM24094T) were randomly selected from the 27 Aeromonas species for experimental validation.Results/key findings. The twelve type strains demonstrated distinctive RFLP patterns and supported the in silico digestion. Subsequently, 60 clinical and environmental strains from our collection, comprising nine Aeromonas species, were used for screening examinations, and the results were in agreement.

CONCLUSION: This method provides an alternative method for laboratory identification, surveillance and epidemiological investigations of clinical and environmental specimens.

Molecular identification and biofilm-forming ability of Elizabethkingia species

Puah SM, Fong SP, Kee BP, Puthucheary SD, Chua KH

Microb Pathog, 2022 Jan;162:105345.

PMID: 34896547 DOI: 10.1016/j.micpath.2021.105345

Recently, Elizabethkingia species have gained attention as a cause of life-threatening infections. The identification via phenotypic methods of three important species- Elizabethkingia meningoseptica, E. anophelis and E. miricola is difficult. Our objectives were to re-assess 30 archived Flavobacterium meningosepticum isolates using 16S rRNA gene sequencing, ERIC-PCR, and biofilm formation assay. Twenty-four isolates were re-identified as E. anophelis and 6 as E. miricola. All of them had the ability to form biofilm as shown in microtiter plate assay based on crystal violet staining. Overall, E. anophelis had a higher specific biofilm formation index compared to E. miricola. A total of 42% (10 out of 24) of E. anophelis were classified as strong, 29% (7 out of 24) as moderate and 29% (7 out of 24) as weak biofilm producers. E. miricola, 17% (1 out of 6) isolates were strong biofilm producers, 50% (3 out of 6) moderate and 33% (2 out of 6) were weak producers. E. anophelis from tracheal secretions were significantly associated with (p = 0.0361) strong biofilm formation. In summary, this study showed that the isolates originally identified as F. meningosepticum were re-classified using the 16S rRNA gene as one of two Elizabethkingia species. The ability of E. anophelis to form strong biofilm in endotracheal tubes indicates their probable role in the pathogenesis of Elizabethkingia infections.

Simple and rapid detection of Salmonella strains by direct PCR amplification of the hilA gene

Pathmanathan SG, Cardona-Castro N, Sánchez-Jiménez MM, Correa-Ochoa MM, Puthucheary SD, Thong KL

J Med Microbiol, 2003 Sep;52(Pt 9):773-6.

PMID: 12909653

The suitability of a PCR procedure using a pair of primers targeting the hilA gene was evaluated as a means of detecting Salmonella species. A total of 33 Salmonella strains from 27 serovars and 15 non-Salmonella strains from eight different genera were included. PCR with all the Salmonella strains produced a 784 bp DNA fragment that was absent from all the non-Salmonella strains tested. The detection limit of the PCR was 100 pg with genomic DNA and 3 x 10(4) c.f.u. ml(-1) with serial dilutions of bacterial culture. An enrichment-PCR method was further developed to test the sensitivity of the hilA primers for the detection of Salmonella in faecal samples spiked with different concentrations of Salmonella choleraesuis subsp. choleraesuis serovar Typhimurium. The method described allowed the detection of Salmonella Typhimurium in faecal samples at a concentration of 3 x 10(2) c.f.u. ml(-1). In conclusion, the hilA primers are specific for Salmonella species and the PCR method presented may be suitable for the detection of Salmonella in faeces.

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