RESEARCH ARTICLE


Molecular Detection and PCR-RFLP Analysis of Mucoviscosity-Associated Gene A (magA) in Clinical Isolates of Multidrug-Resistant Klebsiella pneumoniae in Bangladesh



Md. Hazrat Ali1, *, Saeed Anwar1, Nusrat Jahan Toma1, , Ikram Rafid1, , Md. Kamrul Hasan2, Md. Javed Foysal1, 3
1 Department of Genetic Engineering and Biotechnology, Shahjalal University of Science and Technology, Sylhet 3114, Bangladesh
2 Department of Biochemistry and Molecular Biology, Tejgaon College, National University, Gazipur 1704, Bangladesh
3 School of Molecular and Life Sciences, Curtin University, Kent Street, Bentley, Perth, WA6102, Australia


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Creative Commons License
© 2020 Ali et al.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: https://creativecommons.org/licenses/by/4.0/legalcode. This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.


These authors contributed equally to this work.* Address correspondence to this author at Department of Genetic Engineering and Biotechnology, 211 - Academic Building E, Shahjalal University Avenue, Shahjalal University of Science and Technology, Sylhet 3114, Bangladesh; Tel: +88 01716 299 245; E-mail:hazratsust05@gmail.com


Abstract

Background and Objective:

The mucoviscosity associated gene A (magA) in the hypermucoviscous variants of K. pneumoniae is reported to be associated with invasive infections and considered a virulence factor. We sought to analyze the magA genes in K. pneumoniae isolates in the clinical specimen collected from Bangladesh.

Methods:

We established a multicenter cohort of patients with Klebsiella infection hospitalized at 05 different hospitals between September 2016 and April 2017. We collected 313 K. pneumoniae isolates from patients who consented to participate in the study. The isolates were evaluated for harboring the magA genes using a single-tube multiplexed polymerase chain reaction. The magA genes were analyzed by PCR-RFLP technique using two enzymes, namely PciI and SmaI. Antibiogram assay using 12 commercially available antibiotic discs was performed on all the isolates.

Results:

The presence of K. pneumoniae specific gene (ureD) was confirmed in all the isolates. The percentage of isolates harboring the magA gene was 7.34%(23 isolates), the majority of which was collected from the patients admitted in intensive care units (16 isolates, 69.6%), and infectious diseases wards (5 isolates, 21.7%). PCR-RFLP analysis revealed that for 7 out of 23 isolates, where Sma1 could not cleave the magA gene. All the isolates showed resistance to ampicillin, carbenicillin cefradine, chloramphenicol, erythromycin, kanamycin, and sulphamethoxazole, though the extent was varying. However, imipenem showed 100% sensitivity to all the tested isolates.

Conclusion:

This study demonstrates the presence of the magA gene in multidrug-resistant clinical isolates of K. pneumoniae collected from Bangladesh.

Keywords: Klebsiella pneumoniae, Polysaccharide capsule, ureD, magA, Multidrug-resistance, PCR-RFLP, Imipenem, Antibiogram.