Showing the limitations of available phenotypic assays to detect Burkholderia pseudomallei from clinical specimens in Nigeria

Abstract

The genus Burkholderia comprises Gram-negative bacteria that are metabolically complex and versatile, often thriving in hostile settings. Burkholderia pseudomallei , the causative agent of melioidosis, is a prominent member of the genus and a clinical pathogen in tropical and sub-tropical regions. This pathogen is well known for its multidrug resistance and possible bioweapon potential. There is currently no report of the pathogen from clinical specimens in Nigeria, which might be due to misdiagnosis with phenotypic assays. This study aims to explore the accuracy of the use of phenotypic assays to diagnose B. pseudomallei in Nigeria. Two hundred and seventeen clinical samples and 28 Gram-negative clinical isolates were collected and analysed using Ashdown’s selective agar and monoclonal antibody-based latex agglutination. Species-level identification was achieved using the analytical profile index (API) 20NE system. The susceptibility of the isolates to nine different antimicrobial agents was determined using the disc diffusion method. A total of seventy-four culture-positive isolates were obtained using Ashdown’s selective agar. Twenty-two of these isolates were believed to be B. pseudomallei through the monoclonal antibody-based latex agglutination test and the API 20NE system subsequently identified 14 isolates as Burkholderia . The predominant Burkholderia species was B. cepacia with an isolation rate of 30.8 % (8/26). No isolate was distinctively identified as B. pseudomallei but five isolates were strongly suspected to be B. pseudomallei with similarity indices ranging from 81.9–91.3 %. Other bacterial species with definitive identity include Aeromonas sp., Sphingomonas sp. and Pseudomonas aeruginosa . The antibiotic susceptibility results revealed an overall resistance to amoxicillin–clavullanic acid of 71.4 %, to cefepime of 33.3 %, to trimethoprim–sulfamethoxazole of 38.1 %, to piperacillin–tazobactam of 33.3 %, to imipenem of 66.7 %, to doxycycline of 57.1% and to ceftazidime of 66.7 %. The highest intermediate resistance was observed for cefepime and piperacillin–tazobactam with a value of 66.7 % each, while there was no intermediate resistance for gentamicin, colistin and imipenem. Our findings, therefore, show that phenotypic assays alone are not sufficient in the diagnosis of melioidosis. Additionally, they provide robust support for present and future decisions to expand diagnostic capability for melioidosis beyond phenotypic assays in low-resource settings.