Molecular detection of Vibrio cholerae from human stool collected from SK Hospital, Mymensingh, and their antibiogram.

Farah Zereen,Soudiya Akter,Md Rahman,Md Sobur,Muhammad Hossain

doi:10.5455/javar.2019.f367

Farah Zereen, Soudiya Akter + Show 3 more

Open Access

https://doi.org/10.5455/javar.2019.f367

Copy DOI

Abstract

Objective:Vibrio spp., particularly, Vibrio cholerae is a major etiology of diarrhea in humans worldwide. In this study, we isolated and identified V. cholerae from the human stool of suspected cases along with antibiogram.Materials and Methods:In total, 25 stool samples from cholera suspected patients were analyzed. Isolation and molecular detection of Vibrio species were performed based on staining, motility, cultural and biochemical characteristics followed by polymerase chain reaction (PCR) using groEL gene-specific primers.Results:Among the 25 samples, seven showed growth of yellow color colonies on Thiosulfate-Citrate-Bile salts-Sucrose agar plates. The isolates were Gram-negative, curved shaped, and motile. Biochemically, they were found positive for indole and Methyl Red tests and negative for Voges–Proskauer test. Out of the seven positive samples, only three isolates were confirmed as Vibrio spp. using genus-specific primers. Subsequently, these three isolates were confirmed as V. cholerae by PCR using V. cholerae groEL gene-specific primers. Antibiotic sensitivity test revealed these three isolates as highly sensitive to azithromycin, chloramphenicol, gentamicin, and norfloxacillin while resistant to streptomycin, tetracycline, and oxacillin.Conclusion:Vibrio cholerae were isolated from the stool of diarrheic human patients and confirmed by PCR targeting the groEL gene. The isolates were found resistant to streptomycin, tetracycline and oxacillin, and need further characterization to reveal the molecular basis of their origin and resistance.

Highlights

The genus Vibrio is a member of the family Vibrioneceae
Isolation and molecular detection of Vibrio species were performed based on staining, motility, cultural and biochemical characteristics followed by polymerase chain reaction (PCR) using groEL gene-specific primers
The isolates were found resistant to streptomycin, tetracycline and oxacillin, and need further characterization to reveal the molecular basis of their origin and resistance

Summary

Introduction

The genus Vibrio is a member of the family Vibrioneceae. Vibrio is Gram-negative and porogenous rods having straight or curved rod shape. There are 72 species under this genus, of which 12 species occur in human clinical samples [2] Among these 12 species, V. cholerae, V. parahaemolyticus, and V. vulnificus account for the majority of Vibrio infections in humans [3]. A major proportion of human Vibrio infections are caused due to consumption of contaminated water and foods [12,13]. Vibrio cholera resistance to antibiotics including ampicillin, gentamicin, kanamycin, streptomycin, tetracycline, trimethoprim, and sulfonamides has become a serious treatment problem in many countries of the world, including developing countries like Bangladesh [20]. The present study was set up to isolate and identify V. cholerae from stool samples by conventional bacteriological methods followed by polymerase chain reaction (PCR) with new groEL gene-specific primers that have not been used previously in Bangladesh.

Ethical approval

Results and Discussion

Conclusion

Conflict of Interest