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Title: Evaluation of consensus method for the culture of Burkholderia pseudomallei in soil samples from Laos [version 2; referees: 2 approved]
Authors: David A.B. Dance
Michael Knappik
Sabine Dittrich
Viengmon Davong
Joy Silisouk
Manivanh Vongsouvath
Sayaphet Rattanavong
Alain Pierret
Paul N. Newton
Premjit Amornchai
Vanaporn Wuthiekanun
Sayan Langla
Direk Limmathurotsakul
London School of Hygiene & Tropical Medicine
University of Oxford
Mahidol University
Foundation for Innovative and New Diagnostics (FIND)
UPMC Univ Paris 06
Mahosot Hospital
Médecins Sans Frontières
Keywords: Biochemistry, Genetics and Molecular Biology;Medicine
Issue Date: 1-Jan-2018
Citation: Wellcome Open Research. Vol.3, (2018)
Abstract: © 2018 Dance DAB et al. Background: We have previously shown that PCR following enrichment culture is the most sensitive method to detect Burkholderia pseudomallei in environmental samples. Here we report an evaluation of the published consensus method for the culture of B. pseudomallei from Lao soil in comparison with our conventional culture method and with PCR with or without prior broth enrichment. Methods: One hundred soil samples were collected from a field known to contain B. pseudomallei and processed by: (i) the conventional method, (ii-iii) the consensus method using media prepared in either Laos or Thailand, and (iv) the consensus method performed in Thailand, as well as by (v) PCR following direct extraction of DNA from soil and (vi) PCR following broth pre-enrichment. Results: The numbers of samples in which B. pseudomallei was detected were 42, 10, 7, 6, 6 and 84, respectively. However, two samples were positive by the consensus method but negative by conventional culture, and one sample was negative by PCR following enrichment although B. pseudomallei was isolated by the conventional culture method. Conclusions/Discussion: The results show that no single method will detect all environmental samples that contain B. pseudomallei. People conducting environmental surveys for this organism should be aware of the possibility of false-negative results using the consensus culture method. An approach that entails screening using PCR after enrichment, followed by the evaluation of a range of different culture methods on PCR-positive samples to determine which works best in each setting, is recommended.
ISSN: 2398502X
Appears in Collections:Scopus 2018

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