Survey and Rapid detection of Bordetella pertussis in clinical samples targeting the BP485 in China

Bordetella pertussis is an important human respiratory pathogen. Here, we describe a loop-mediated isothermal amplification (LAMP) method for the rapid detection of B. pertussis in clinical samples based on a visual test. The LAMP assay detected the BP485 target sequence within 60 min with a detecti...

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Main Authors: Wei eLiu, Hua Ying Xu, Rong De Dong, Huan eLi, Na Xiang Zhao, Li Li Li, Ying eZhang, Xiao eWei, Song Xue Wang, Mo Si Huang, Ming eZeng, Yu Liu Huang, Min Shu Zhang, Jing eYuan
Format: Article
Language:English
Published: Frontiers Media S.A. 2015-03-01
Series:Frontiers in Public Health
Subjects:
Online Access:http://journal.frontiersin.org/Journal/10.3389/fpubh.2015.00039/full
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author Wei eLiu
Hua Ying Xu
Rong De Dong
Huan eLi
Na Xiang Zhao
Li Li Li
Ying eZhang
Xiao eWei
Song Xue Wang
Mo Si Huang
Ming eZeng
Yu Liu Huang
Min Shu Zhang
Jing eYuan
author_facet Wei eLiu
Hua Ying Xu
Rong De Dong
Huan eLi
Na Xiang Zhao
Li Li Li
Ying eZhang
Xiao eWei
Song Xue Wang
Mo Si Huang
Ming eZeng
Yu Liu Huang
Min Shu Zhang
Jing eYuan
author_sort Wei eLiu
collection DOAJ
description Bordetella pertussis is an important human respiratory pathogen. Here, we describe a loop-mediated isothermal amplification (LAMP) method for the rapid detection of B. pertussis in clinical samples based on a visual test. The LAMP assay detected the BP485 target sequence within 60 min with a detection limit of 1.3 pg/µl, a 10-fold increase in sensitivity compared with conventional PCR. All 31 non-pertussis respiratory pathogens tested were negative for LAMP detection, indicating the high specificity of the primers for B. pertussis. To evaluate the application of the LAMP assay to clinical diagnosis, of 105 sputum and nasopharyngeal samples collected from the patients with suspected respiratory infections in China, a total of 12 Bordetella pertussis isolates were identified from 33 positive samples detected by LAMP-based surveillance targeting BP485. Strikingly, a 4.5 months old baby and her mother were found to be infected with B. pertussis at the same time. All isolates belonged to different B. pertussis multilocus sequence typing (MLST) groups with different alleles of the virulence-related genes including 4 alleles of ptxA, 6 of prn, 4 of tcfA, 2 of fim2 and 3 of fim3. The diversity of B. pertussis carrying toxin genes in clinical strains indicates a rapid and continuing evolution of B. pertussis. This combined with its high prevalence will make it difficult to control. In conclusion, we have developed a visual detection LAMP assay, which could be a useful tool for rapid B. pertussis detection, especially in situations where resources are poor and in point-of-care tests.
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spelling doaj.art-72f23999c1634d66af71e4a4b9a62e212022-12-22T00:11:29ZengFrontiers Media S.A.Frontiers in Public Health2296-25652015-03-01310.3389/fpubh.2015.00039128239Survey and Rapid detection of Bordetella pertussis in clinical samples targeting the BP485 in ChinaWei eLiu0Hua Ying Xu1Rong De Dong2Huan eLi3Na Xiang Zhao4Li Li Li5Ying eZhang6Xiao eWei7Song Xue Wang8Mo Si Huang9Ming eZeng10Yu Liu Huang11Min Shu Zhang12Jing eYuan13Institute of Disease Control and Prevention,Academy of Military Medical SciencesNational Institutes for Food and Drug ControlInstitute of Disease Control and Prevention,Academy of Military Medical SciencesInstitute of Disease Control and Prevention,Academy of Military Medical SciencesInstitute of Disease Control and Prevention,Academy of Military Medical SciencesNational Institutes for Food and Drug ControlNational Institutes for Food and Drug ControlInstitute of Disease Control and Prevention,Academy of Military Medical SciencesInstitute of Disease Control and Prevention,Academy of Military Medical SciencesInstitute of Disease Control and Prevention,Academy of Military Medical SciencesNational Institutes for Food and Drug ControlInstitute of Disease Control and Prevention,Academy of Military Medical SciencesNational Institutes for Food and Drug ControlInstitute of Disease Control and Prevention,Academy of Military Medical SciencesBordetella pertussis is an important human respiratory pathogen. Here, we describe a loop-mediated isothermal amplification (LAMP) method for the rapid detection of B. pertussis in clinical samples based on a visual test. The LAMP assay detected the BP485 target sequence within 60 min with a detection limit of 1.3 pg/µl, a 10-fold increase in sensitivity compared with conventional PCR. All 31 non-pertussis respiratory pathogens tested were negative for LAMP detection, indicating the high specificity of the primers for B. pertussis. To evaluate the application of the LAMP assay to clinical diagnosis, of 105 sputum and nasopharyngeal samples collected from the patients with suspected respiratory infections in China, a total of 12 Bordetella pertussis isolates were identified from 33 positive samples detected by LAMP-based surveillance targeting BP485. Strikingly, a 4.5 months old baby and her mother were found to be infected with B. pertussis at the same time. All isolates belonged to different B. pertussis multilocus sequence typing (MLST) groups with different alleles of the virulence-related genes including 4 alleles of ptxA, 6 of prn, 4 of tcfA, 2 of fim2 and 3 of fim3. The diversity of B. pertussis carrying toxin genes in clinical strains indicates a rapid and continuing evolution of B. pertussis. This combined with its high prevalence will make it difficult to control. In conclusion, we have developed a visual detection LAMP assay, which could be a useful tool for rapid B. pertussis detection, especially in situations where resources are poor and in point-of-care tests.http://journal.frontiersin.org/Journal/10.3389/fpubh.2015.00039/fullPrevalencespecificityLAMPSensitivityrapid diagnosisB. pertussis
spellingShingle Wei eLiu
Hua Ying Xu
Rong De Dong
Huan eLi
Na Xiang Zhao
Li Li Li
Ying eZhang
Xiao eWei
Song Xue Wang
Mo Si Huang
Ming eZeng
Yu Liu Huang
Min Shu Zhang
Jing eYuan
Survey and Rapid detection of Bordetella pertussis in clinical samples targeting the BP485 in China
Frontiers in Public Health
Prevalence
specificity
LAMP
Sensitivity
rapid diagnosis
B. pertussis
title Survey and Rapid detection of Bordetella pertussis in clinical samples targeting the BP485 in China
title_full Survey and Rapid detection of Bordetella pertussis in clinical samples targeting the BP485 in China
title_fullStr Survey and Rapid detection of Bordetella pertussis in clinical samples targeting the BP485 in China
title_full_unstemmed Survey and Rapid detection of Bordetella pertussis in clinical samples targeting the BP485 in China
title_short Survey and Rapid detection of Bordetella pertussis in clinical samples targeting the BP485 in China
title_sort survey and rapid detection of bordetella pertussis in clinical samples targeting the bp485 in china
topic Prevalence
specificity
LAMP
Sensitivity
rapid diagnosis
B. pertussis
url http://journal.frontiersin.org/Journal/10.3389/fpubh.2015.00039/full
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