Multiple ambler class A ESBL genes among Klebsiella pneumoniae isolates in a Malaysian district hospital.

Detailed reports regarding the distribution and activity of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae isolates are currently not widely available in the Malaysian setting. This study was conducted to determine the ESBL genes distribution rate, phenotypic detection, and antimicrobial susceptibility patterns among betalactam resistant Klebsiella pneumoniae isolated from a Malaysian district hospital. K. pneumoniae isolates were collected from a microbiology laboratory at Hospital Pakar Sultanah Fatimah, Malaysia. Following exclusion and inclusion criteria, 141 isolates were selected for this study. K. pneumoniae was identified by phenotypic method, whilst antibiotics' susceptibility patterns were determined by the Kirby-Bauer method, as described in Clinical Laboratory Standard Institute (CLSI) guidelines (Oxoid, UK; Becton-Dickenson, USA). Detection of Ambler Group A ESBL gene (blaSHV, blaTEM, blaCTX-M-1, blaCTX-M-2, blaCTX-M-8, blaCTX-M-9, and blaCTX-M-25) was done using polymerase chain reaction (PCR). ESBL genes were found in 85.8% of K. pneumoniae (121 of 141) isolates. Only blaSHV, blaTEM, blaCTX-M-1, and blaCTX-M-9 were detected among K. pneumoniae isolates with distribution rates of 75.2% (106 of 141), 41.1% (58 of 141), 44% (62 of 141), and 0.7% (1 of 141), respectively. There was no blaCTX-M-2, blaCTX-M-8, or blaCTX-M-25 detected from any isolates in this study. Sequencing of representative amplicons revealed blaSHV as SHV-12, blaTEM as TEM-1, blaCTX-M-1 as CTX-M-15, and blaCTX-M-9 as CTX-M-18. The phenotypic detection rate of ESBL was 71.6% (101 of 141), whilst 9.2% (13 of 141) were positive for carbapenemase. AmpC betalactamase was detected in 22% (31 of 141) of all isolates. Antibiotic resistance was between 44.6% (netilmicin) and 97.2% (cefotaxime). Based on ESBL genes distribution, blaSHV was a predominant gene found in one of Malaysian district hospitals, notwithstanding having blaTEM, blaCTX-M-1, and blaCTX-M-9. Despite carrying multiple ESBL genes, some strains were positive for carbapenemase or AmpC betalactamase, which resulted in high antimicrobial resistance rates.

Multiple ambler class A ESBL genes among Klebsiella pneumoniae isolates in a Malaysian district hospital

Detailed reports regarding the distribution and activity of extended-spectrum beta-lactamase (ESBL)-producing Klebsiella pneumoniae isolates are currently not widely available in the Malaysian setting. This study was conducted to determine the ESBL genes distribution rate, phenotypic detection, and antimicrobial susceptibility patterns among betalactam resistant Klebsiella pneumoniae isolated from a Malaysian district hospital. K. pneumoniae isolates were collected from a microbiology laboratory at Hospital Pakar Sultanah Fatimah, Malaysia. Following exclusion and inclusion criteria, 141 isolates were selected for this study. K. pneumoniae was identified by phenotypic method, whilst antibiotics’ susceptibility patterns were determined by the Kirby-Bauer method, as described in Clinical Laboratory Standard Institute (CLSI) guidelines (Oxoid, UK; Becton-Dickenson, USA). Detection of Ambler Group A ESBL gene (blaSHV, blaTEM, blaCTX-M-1, blaCTX-M-2, blaCTX-M-8, blaCTX-M-9, and blaCTX-M-25) was done using polymerase chain reaction (PCR). ESBL genes were found in 85.8% of K. pneumoniae (121 of 141) isolates. Only blaSHV, blaTEM, blaCTX-M-1, and blaCTX-M-9 were detected among K. pneumoniae isolates with distribution rates of 75.2% (106 of 141), 41.1% (58 of 141), 44% (62 of 141), and 0.7% (1 of 141), respectively. There was no blaCTX-M-2, blaCTX-M-8, or blaCTX-M-25 detected from any isolates in this study. Sequencing of representative amplicons revealed blaSHV as SHV-12, blaTEM as TEM-1, blaCTX-M-1 as CTX-M-15, and blaCTX-M-9 as CTX-M-18. The phenotypic detection rate of ESBL was 71.6% (101 of 141), whilst 9.2% (13 of 141) were positive for carbapenemase. AmpC betalactamase was detected in 22% (31 of 141) of all isolates. Antibiotic resistance was between 44.6% (netilmicin) and 97.2% (cefotaxime). Based on ESBL genes distribution, blaSHV was a predominant gene found in one of Malaysian district hospitals, notwithstanding having blaTEM, blaCTX-M-1, and blaCTX-M-9. Despite carrying multiple ESBL genes, some strains were positive for carbapenemase or AmpC betalactamase, which resulted in high antimicrobial resistance rates.

Evaluation of PCR-based approach for serotype determination of Streptococcus pneumoniae.

Determination of Streptococcus pneumoniae serotypes is essential for epidemiological surveillance. Therefore accurate, reliable and cost effective serotyping method is crucial. In this study, we determined the serotypes of 41 pneumococcal isolates recovered from human anterior nares by multiplex Polymerase Chain Reaction (PCR) utilizing published primers. The data was then compared with conventional serology using latex agglutination (LA) and the Quellung reaction. Based on the PCR-approach, 8 different serogroups/serotypes were detected with one isolate classified as non-typeable (cpsA-negative). In reference to the serology-based data, the results were in agreement except for one isolate. For the latter isolate, the LA and Quellung tests failed to show a reaction but the PCR-approach and sequencing identified the isolate as serogroup 15B/C. Based on this experimental setting, we found that the PCR-approach for pneumococcal serotypes determination is reliable to serve as the alternative for determining the pneumococcal serotyping.

Testicular relapse in childhood acute lymphoblastic leukaemia in Malaysia, 1967-82.

From 1967-82, 9 children with testicular relapse (TR) of acute lymphoblastic leukaemia (ALL) were diagnosed out of 99 boys treated, an incidence of 9.1%. The median time from the onset of ALL until diagnosis was 28 months (range 3-41 months). All were asymptomatic; six were detected on routine examination while three were diagnosed only on biopsy. Routine biopsy prior to stopping chemotherapy is useful in detecting occult TR. Biopsies should be done on both the testes regardless of the clinical findings. The age, leucocyte count and hepatosplenomegaly at diagnosis of ALL were not found to be significant factors in influencing relapse. Eight children were in bone marrow remission at the time of TR, but three had preceding or concurrent meningeal leukaemia while in the other five the testis was the first and only site of relapse. Radiotherapy was effective in local disease control but failed to prevent bone marrow relapse in all except two patients despite continuation of chemotherapy. The median time from onset of TR until bone marrow relapse was 7 months (range 3-13 months) and the median time until death, was 11 months (range 6-18 months). The frequency of testicular relapse may be related to the intensity of either the initial induction therapy or the consolidation chemotherapy. Further studies are required to determine whether the incidence of testicular relapse will decline with more intensive early treatment.