Background Carbapenem-resistant Enterobacteriaceae (CRE) are the most critical group of MDR bacteria that pose a threat to human health especially affecting patients with haematological malignancies. Knowledge on the prevalence of CRE colonization of the gut will help healthcare providers to be vigilant in-patient management. However, there are no local studies exploring the gut colonizing CRE among patients with haematological malignancies. Hence, this study aimed to determine the prevalence of CRE genes in colonizing CRE isolates in patients with haematological malignancies and evaluate their antibiotic susceptibility pattern. Methods Rectal swab samples collected from patients with haematological malignancies were screened with selective chromogenic agar for CRE. Species identification and ABST of CRE isolates were done using VITEK® 2 system. blaKPC, blaNDM and blaOXA−48 genes were identified using an in-house conventional multiplex PCR, with previously described method and primers. Results A total of 264 adult patients with haematological malignancies were included in the study. Prevalence of gut colonization of CRE was 35.2% (93/264). A total of 119 CRE isolates from 93 study subjects were further studied. Klebsiella pneumoniae was the predominant carbapenemase producer [68/119 (57%)] among the isolates. Sixty-one (51.2%) isolates harboured blaNDM, 48 (40.3%) blaOXA−48 and 12 (10%) blaKPC. Three isolates (2.5%) were shown to harbour all three genes and 24 (20%) two genes. Twenty (16.8%) of isolates tested negative for all three genes. Conclusions A relatively high prevalence of CRE gene occurrence and co-occurrence was detected in the study population. This indicates a potential challenge to infection prevention and control in the institute as colonization with CRE carries a threat to endogenous infection and cross transfer. Limitations In-house conventional PCR method was used to identify CRE genes. Twenty isolates that were negative for all tested genes need to be studied further.
Background Carbapenemase-producing Enterobacteriaceae (CRE) are a global problem and prevalence of CR genes differs across geographical regions. The current Sri Lankan situation is unknown; hence, this study aimed to identify three commonly occurring CR genes in the region. Methods Enterobacteriaceae isolates resistant to carbapenems were collected from clinical samples submitted for routine testing to the microbiology laboratory of National Cancer Institute. Species identification was done using conventional biochemicals and CLSI disc diffusion method was used for ABST of CRE isolates. Identification of CR genes was done by in-house conventional multiplex PCR, using previously described primers (Table 1). Ten microlitres of total DNA from each sample, extracted by heat shock method, was subjected to multiplex PCR in a 50 μL of PCR mixture which contained 5× PCR buffer (10 mmol/L Tris–HCl, 50 mmol/L KCl), 2.5 mmol/L of MgCl2, 10 mmol/L of deoxynucleotide triphosphate, 5 U/mL of Taq Polymerase and 20 μmol/L of each primer. The PCR program was run at 94°C for 3 min, followed by 30 cycles of 95°C for 1 min, 55°C for 30 s and 73°C for 1 min, with a final extension at 72°C for 5 min. Products were visualized by electrophoresis in 2% agarose gel (Figure 1). Results A total of 123 isolates were tested. Klebsiella pneumoniae was the predominant carbapenemase producer (58.5%). Sixty-six (53.6%) isolates harboured blaOXA-48 and 47 (38.2%) had blaNDM. Only one isolate was positive for blaKPC. Thirteen (10.5%) had both blaOXA-48 and blaNDM and 9 (7%) tested negative for all three genes. Conclusions bla OXA-48 and blaNDM being common and blaKPC being very rare is expected with the geographical location of the country. Noted high prevalence and co-occurrence of CR genes among CRE isolates is alarming and pose a threat to patient management and infection control of the institution.
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