Development of a PCR-dipstick DNA chromatography-based tool for the detection of CTX-M- and TEM-producing Escherichia coli and Klebsiella pneumoniae isolated from patients in Kafue and Katete districts of Zambia
Rattachement africain : jp, Zambie, us. Niveau de preuve : code pays fourni par la source.
Le résumé fourni par la source
In Zambia, 40% of clinical Gram-negative bacteria are either Escherichia coli or Klebsiella pneumoniae , with a high third-generation cephalosporin (3GC) resistance prevalence. Therefore, 3GC resistance surveillance is a crucial indicator for guiding focused intervention policies. However, the lack of genotypic diagnostic tools limits the ability to elucidate trends, especially in peri-urban and rural areas of developing countries. This study aimed to develop a rapid, cost-effective tool for the genotypic surveillance of 3GC resistance. Here, 900 stool samples collected from patients in Kafue (peri-urban, n = 400) and Katete (rural, n = 500) districts of Zambia were used for bacterial isolation on MacConkey agar supplemented with 1 μg/ml cefotaxime. Isolated 3GC-resistant strains were characterized by sequencing the 16S rRNA gene and screening for bla CTX-M and bla TEM genes using single polymerase chain reaction (PCR). Furthermore, selected 3GC-resistant strains were subjected to whole-genome sequencing (WGS) using MiSeq/HiSeq ( n = 34) and MinION ( n = 1). Using the data from this and other previous studies, we developed a rapid PCR-dipstick DNA chromatography-based tool for detecting bla CTX-M , bla TEM , E. coli -specific yaiO, and K. pneumoniae -specific khe genes. The prevalence of isolated 3GC resistant strains was 15.4% (139/900), dominated by E. coli (102/139, 73.4%). On PCR, the bla CTX-M gene was detected in 72.7% (101/139) of the isolates, while bla TEM was found in 46.8% (65/139) of the strains. The developed tool displayed a high level of agreement with WGS and single PCR/Sanger sequencing, with sensitivity and specificity ≥ 95% and Kappa ≥ 0.95 for each of the four target genes. We envisage that the simplicity and adaptability of this tool will be a significant advantage for the surveillance of 3GC resistance in Zambia and elsewhere.
Ce résumé expose les affirmations des auteurs. BNTIC ne l’interprète pas comme une validation indépendante des résultats.
Le contrôle bibliographique ouvert
DOI retrouvé dans Crossref DOI retrouvé ; titre concordant.
- Titre Crossref
- Development of a PCR-dipstick DNA chromatography-based tool for the detection of CTX-M- and TEM-producing Escherichia coli and Klebsiella pneumoniae isolated from patients in Kafue and Katete districts of Zambia
- Date Crossref
- 16/04/2025
- Éditeur
- Springer Science and Business Media LLC
- Type
- journal-article
Ce recoupement confirme des métadonnées liées au DOI. Il ne confirme ni la méthode ni les conclusions de l’étude, et il ne compte pas comme une seconde source scientifique indépendante.
Où se fait cette recherche
-
Hokkaido University International Institute for Zoonosis Control Lusaka, Zambie (pays nommé en fin d’affiliation)Université ou école supérieure
-
University of Zambia Zambie (code pays fourni par la source)Université ou école supérieure
-
Ministry of Agriculture and Livestock Zambie (code pays fourni par la source)Organisme public
-
Lusaka Apex Medical University Lusaka Apex Medical University, Zambie (code pays fourni par la source)Université ou école supérieure
-
Levy Mwanawasa Medical University Institute of Basic and Biomedical Sciences Lusaka, Zambie (code pays fourni par la source)Université ou école supérieure
-
National Institute of Infectious Diseases Department of Virology-I pays non établi dans la noticeOrganisme public
-
University Teaching Hospital Department of Pathology and Microbiology Lusaka, Zambie (code pays fourni par la source)Établissement de santé
-
Ministry of Health Zambie (code pays fourni par la source)Organisme public
-
Hokkaido University of Science pays non établi dans la noticeUniversité ou école supérieure
-
Centre for Infectious Disease Research in Zambia Zambie (code pays fourni par la source)Organisation à but non lucratif
-
Global Virus Network pays non établi dans la noticeInstitution
-
GenEndeavor LLC pays non établi dans la noticeInstitution
International Institute for Zoonosis Control — Hokkaido University (Lusaka, Zambie), University of Zambia (Zambie) et Ministry of Agriculture and Livestock (Zambie), avec 9 autres affiliations. Pays d’affiliation : Zambie.
Une affiliation ne permet pas de déduire la nationalité d’un auteur.