Detection of Wolbachia in wild mosquito populations from selected areas in Peninsular Malaysia by loop-mediated isothermal amplification (LAMP) technique

M. M. Noor-Shazleen-Husnie, Emelia Osman, M. S. Ahmad-Firdaus, P. Zainol-Ariffin, Aishah Hani Azil

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Wolbachia, a naturally endosymbiotic bacteria, has shown its potential as one of biological control agents for vector borne diseases. Due to large number of mosquitoes involved in Wolbachia screening, Loop-mediated isothermal amplification (LAMP) is recommended as a convenient and time-saving technique. This study aimed to evaluate a newly developed LAMP assay for detection of Wolbachia by targeting 16S rDNA gene in samples of wild mosquito populations. The LAMP products were confirmed by colorimetric detection using hydroxy naphthol blue (HNB), digestion with RsaI restriction enzyme and gel electrophoresis. The restriction enzyme digestion of PCR products was performed to differentiate between Wolbachia supergroups A and B. Out of 765 mosquito samples tested, 349 (45.6%) and 237 (31%) of the samples were positive for LAMP and PCR techniques respectively. The prevalence of Wolbachia detected in mosquitoes was significantly higher using LAMP as compared to PCR. There is significant association between numbers of mosquitoes positive with Wolbachia detected using LAMP and PCR (χ2 =61.31; df=1; p < 0.05) with a kappa (κ) value of 0.27 and Phi value, 0.283. This study highlighted the potential of LAMP as a sensitive, specific and rapid tool for screening of Wolbachia in mosquitoes, thus it presents as an alternative to PCR-based assays.

Original languageEnglish
Pages (from-to)330-346
Number of pages17
JournalTropical Biomedicine
Volume35
Issue number2
Publication statusPublished - 1 Jun 2018

Fingerprint

Wolbachia
Malaysia
Culicidae
Population
Polymerase Chain Reaction
Digestion
Biological Control Agents
Disease Vectors
Enzymes
Ribosomal DNA
Electrophoresis
Gels
Bacteria

ASJC Scopus subject areas

  • Parasitology
  • Infectious Diseases

Cite this

Detection of Wolbachia in wild mosquito populations from selected areas in Peninsular Malaysia by loop-mediated isothermal amplification (LAMP) technique. / Noor-Shazleen-Husnie, M. M.; Osman, Emelia; Ahmad-Firdaus, M. S.; Zainol-Ariffin, P.; Azil, Aishah Hani.

In: Tropical Biomedicine, Vol. 35, No. 2, 01.06.2018, p. 330-346.

Research output: Contribution to journalArticle

@article{5155b95e5a644ca3b551d4e91196f9d6,
title = "Detection of Wolbachia in wild mosquito populations from selected areas in Peninsular Malaysia by loop-mediated isothermal amplification (LAMP) technique",
abstract = "Wolbachia, a naturally endosymbiotic bacteria, has shown its potential as one of biological control agents for vector borne diseases. Due to large number of mosquitoes involved in Wolbachia screening, Loop-mediated isothermal amplification (LAMP) is recommended as a convenient and time-saving technique. This study aimed to evaluate a newly developed LAMP assay for detection of Wolbachia by targeting 16S rDNA gene in samples of wild mosquito populations. The LAMP products were confirmed by colorimetric detection using hydroxy naphthol blue (HNB), digestion with RsaI restriction enzyme and gel electrophoresis. The restriction enzyme digestion of PCR products was performed to differentiate between Wolbachia supergroups A and B. Out of 765 mosquito samples tested, 349 (45.6{\%}) and 237 (31{\%}) of the samples were positive for LAMP and PCR techniques respectively. The prevalence of Wolbachia detected in mosquitoes was significantly higher using LAMP as compared to PCR. There is significant association between numbers of mosquitoes positive with Wolbachia detected using LAMP and PCR (χ2 =61.31; df=1; p < 0.05) with a kappa (κ) value of 0.27 and Phi value, 0.283. This study highlighted the potential of LAMP as a sensitive, specific and rapid tool for screening of Wolbachia in mosquitoes, thus it presents as an alternative to PCR-based assays.",
author = "Noor-Shazleen-Husnie, {M. M.} and Emelia Osman and Ahmad-Firdaus, {M. S.} and P. Zainol-Ariffin and Azil, {Aishah Hani}",
year = "2018",
month = "6",
day = "1",
language = "English",
volume = "35",
pages = "330--346",
journal = "Tropical Biomedicine",
issn = "0127-5720",
publisher = "Malaysian Society of Parasitology and Tropical Medicine",
number = "2",

}

TY - JOUR

T1 - Detection of Wolbachia in wild mosquito populations from selected areas in Peninsular Malaysia by loop-mediated isothermal amplification (LAMP) technique

AU - Noor-Shazleen-Husnie, M. M.

AU - Osman, Emelia

AU - Ahmad-Firdaus, M. S.

AU - Zainol-Ariffin, P.

AU - Azil, Aishah Hani

PY - 2018/6/1

Y1 - 2018/6/1

N2 - Wolbachia, a naturally endosymbiotic bacteria, has shown its potential as one of biological control agents for vector borne diseases. Due to large number of mosquitoes involved in Wolbachia screening, Loop-mediated isothermal amplification (LAMP) is recommended as a convenient and time-saving technique. This study aimed to evaluate a newly developed LAMP assay for detection of Wolbachia by targeting 16S rDNA gene in samples of wild mosquito populations. The LAMP products were confirmed by colorimetric detection using hydroxy naphthol blue (HNB), digestion with RsaI restriction enzyme and gel electrophoresis. The restriction enzyme digestion of PCR products was performed to differentiate between Wolbachia supergroups A and B. Out of 765 mosquito samples tested, 349 (45.6%) and 237 (31%) of the samples were positive for LAMP and PCR techniques respectively. The prevalence of Wolbachia detected in mosquitoes was significantly higher using LAMP as compared to PCR. There is significant association between numbers of mosquitoes positive with Wolbachia detected using LAMP and PCR (χ2 =61.31; df=1; p < 0.05) with a kappa (κ) value of 0.27 and Phi value, 0.283. This study highlighted the potential of LAMP as a sensitive, specific and rapid tool for screening of Wolbachia in mosquitoes, thus it presents as an alternative to PCR-based assays.

AB - Wolbachia, a naturally endosymbiotic bacteria, has shown its potential as one of biological control agents for vector borne diseases. Due to large number of mosquitoes involved in Wolbachia screening, Loop-mediated isothermal amplification (LAMP) is recommended as a convenient and time-saving technique. This study aimed to evaluate a newly developed LAMP assay for detection of Wolbachia by targeting 16S rDNA gene in samples of wild mosquito populations. The LAMP products were confirmed by colorimetric detection using hydroxy naphthol blue (HNB), digestion with RsaI restriction enzyme and gel electrophoresis. The restriction enzyme digestion of PCR products was performed to differentiate between Wolbachia supergroups A and B. Out of 765 mosquito samples tested, 349 (45.6%) and 237 (31%) of the samples were positive for LAMP and PCR techniques respectively. The prevalence of Wolbachia detected in mosquitoes was significantly higher using LAMP as compared to PCR. There is significant association between numbers of mosquitoes positive with Wolbachia detected using LAMP and PCR (χ2 =61.31; df=1; p < 0.05) with a kappa (κ) value of 0.27 and Phi value, 0.283. This study highlighted the potential of LAMP as a sensitive, specific and rapid tool for screening of Wolbachia in mosquitoes, thus it presents as an alternative to PCR-based assays.

UR - http://www.scopus.com/inward/record.url?scp=85052117087&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85052117087&partnerID=8YFLogxK

M3 - Article

AN - SCOPUS:85052117087

VL - 35

SP - 330

EP - 346

JO - Tropical Biomedicine

JF - Tropical Biomedicine

SN - 0127-5720

IS - 2

ER -