Touchdown-touchup nested PCR for low-copy gene detection of benzimidazole-susceptible Wuchereria bancrofti with a Wolbachia endosymbiont imported by migrant carriers.
Identifieur interne : 004A32 ( Main/Exploration ); précédent : 004A31; suivant : 004A33Touchdown-touchup nested PCR for low-copy gene detection of benzimidazole-susceptible Wuchereria bancrofti with a Wolbachia endosymbiont imported by migrant carriers.
Auteurs : Prapassorn Pechgit [Thaïlande] ; Apiradee Intarapuk ; Danai Pinyoowong ; Adisak BhumiratanaSource :
- Experimental parasitology [ 1090-2449 ] ; 2011.
Descripteurs français
- KwdFr :
- ADN bactérien (sang), Adulte, Algorithmes, Animaux, Antihelminthiques (pharmacologie), Benzimidazoles (pharmacologie), Femelle, Filariose lymphatique (diagnostic), Filariose lymphatique (ethnologie), Filariose lymphatique (transmission), Humains, Jeune adulte, Myanmar (ethnologie), Mâle, Population de passage et migrants, Porteur sain (diagnostic), Porteur sain (ethnologie), Porteur sain (parasitologie), Porteur sain (transmission), Réaction de polymérisation en chaîne, Symbiose, Thaïlande, Wolbachia (physiologie), Wuchereria bancrofti (), Wuchereria bancrofti (génétique), Wuchereria bancrofti (isolement et purification), Wuchereria bancrofti (microbiologie).
- MESH :
- diagnostic : Filariose lymphatique, Porteur sain.
- ethnologie : Filariose lymphatique, Myanmar, Porteur sain.
- génétique : Wuchereria bancrofti.
- isolement et purification : Wuchereria bancrofti.
- microbiologie : Wuchereria bancrofti.
- parasitologie : Porteur sain.
- pharmacologie : Antihelminthiques, Benzimidazoles.
- physiologie : Wolbachia.
- sang : ADN bactérien.
- Adulte, Algorithmes, Animaux, Femelle, Filariose lymphatique, Humains, Jeune adulte, Mâle, Population de passage et migrants, Porteur sain, Réaction de polymérisation en chaîne, Symbiose, Thaïlande, Wuchereria bancrofti.
- Wicri :
- geographic : Birmanie.
English descriptors
- KwdEn :
- Adult, Algorithms, Animals, Anthelmintics (pharmacology), Benzimidazoles (pharmacology), Carrier State (diagnosis), Carrier State (ethnology), Carrier State (parasitology), Carrier State (transmission), DNA, Bacterial (blood), Elephantiasis, Filarial (diagnosis), Elephantiasis, Filarial (ethnology), Elephantiasis, Filarial (transmission), Female, Humans, Male, Myanmar (ethnology), Polymerase Chain Reaction, Symbiosis, Thailand, Transients and Migrants, Wolbachia (physiology), Wuchereria bancrofti (drug effects), Wuchereria bancrofti (genetics), Wuchereria bancrofti (isolation & purification), Wuchereria bancrofti (microbiology), Young Adult.
- MESH :
- chemical , blood : DNA, Bacterial.
- chemical , pharmacology : Anthelmintics, Benzimidazoles.
- geographic , ethnology : Myanmar.
- diagnosis : Carrier State, Elephantiasis, Filarial.
- drug effects : Wuchereria bancrofti.
- ethnology : Carrier State, Elephantiasis, Filarial.
- genetics : Wuchereria bancrofti.
- isolation & purification : Wuchereria bancrofti.
- microbiology : Wuchereria bancrofti.
- parasitology : Carrier State.
- physiology : Wolbachia.
- transmission : Carrier State, Elephantiasis, Filarial.
- Adult, Algorithms, Animals, Female, Humans, Male, Polymerase Chain Reaction, Symbiosis, Thailand, Transients and Migrants, Young Adult.
Abstract
A novel, sensitive and specific touchdown-touchup nested PCR (TNPCR) technique based on two useful molecular markers, a Wuchereria bancrofti β-tubulin gene involved in benzimidazole susceptibility and a Wolbachia ftsZ gene involved in cell division, was developed to simultaneously detect the parasite W. bancrofti (W1) with its Wolbachia endosymbiont (W2) from both microfilaremic and post-treatment samples of at-risk migrant carriers infected with geographical W. bancrofti isolates. The detection and characterization of authentically low-copy gene-derived amplicons revealed no false positive identifications in amicrofilaremia with or without antigenemia. The W1-TNPCR was 100-fold more sensitive than the W2-TNPCR regardless of the microfilarial DNA isolation method and compared well with the thick blood film and membrane filtration techniques. These locus-specific TNPCRs could also detect Wolbachia-carrying W. bancrofti genotype in addition to a link to benzimidazole sensitivity among those with unknown infection origins that exhibited microfilaremia responsiveness against treatment with diethylcarbamazine plus albendazole. These TNPCR methods can augment the results of microscopic detection of the parasite because these methods enhance DNA isolation and PCR amplification capabilities.
DOI: 10.1016/j.exppara.2010.10.022
PubMed: 21074526
Affiliations:
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Le document en format XML
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(sang)</term><term>Adulte</term><term>Algorithmes</term><term>Animaux</term><term>Antihelminthiques (pharmacologie)</term><term>Benzimidazoles (pharmacologie)</term><term>Femelle</term><term>Filariose lymphatique (diagnostic)</term><term>Filariose lymphatique (ethnologie)</term><term>Filariose lymphatique (transmission)</term><term>Humains</term><term>Jeune adulte</term><term>Myanmar (ethnologie)</term><term>Mâle</term><term>Population de passage et migrants</term><term>Porteur sain (diagnostic)</term><term>Porteur sain (ethnologie)</term><term>Porteur sain (parasitologie)</term><term>Porteur sain (transmission)</term><term>Réaction de polymérisation en chaîne</term><term>Symbiose</term><term>Thaïlande</term><term>Wolbachia (physiologie)</term><term>Wuchereria bancrofti ()</term><term>Wuchereria bancrofti (génétique)</term><term>Wuchereria bancrofti (isolement et purification)</term><term>Wuchereria bancrofti (microbiologie)</term></keywords><keywords scheme="MESH" type="chemical" 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sain</term><term>Réaction de polymérisation en chaîne</term><term>Symbiose</term><term>Thaïlande</term><term>Wuchereria bancrofti</term></keywords><keywords scheme="Wicri" type="geographic" xml:lang="fr"><term>Birmanie</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">A novel, sensitive and specific touchdown-touchup nested PCR (TNPCR) technique based on two useful molecular markers, a Wuchereria bancrofti β-tubulin gene involved in benzimidazole susceptibility and a Wolbachia ftsZ gene involved in cell division, was developed to simultaneously detect the parasite W. bancrofti (W1) with its Wolbachia endosymbiont (W2) from both microfilaremic and post-treatment samples of at-risk migrant carriers infected with geographical W. bancrofti isolates. The detection and characterization of authentically low-copy gene-derived amplicons revealed no false positive identifications in amicrofilaremia with or without antigenemia. The W1-TNPCR was 100-fold more sensitive than the W2-TNPCR regardless of the microfilarial DNA isolation method and compared well with the thick blood film and membrane filtration techniques. These locus-specific TNPCRs could also detect Wolbachia-carrying W. bancrofti genotype in addition to a link to benzimidazole sensitivity among those with unknown infection origins that exhibited microfilaremia responsiveness against treatment with diethylcarbamazine plus albendazole. These TNPCR methods can augment the results of microscopic detection of the parasite because these methods enhance DNA isolation and PCR amplification capabilities.</div></front></TEI><affiliations><list><country><li>Thaïlande</li></country></list><tree><noCountry><name sortKey="Bhumiratana, Adisak" sort="Bhumiratana, Adisak" uniqKey="Bhumiratana A" first="Adisak" last="Bhumiratana">Adisak Bhumiratana</name><name sortKey="Intarapuk, Apiradee" sort="Intarapuk, Apiradee" uniqKey="Intarapuk A" first="Apiradee" last="Intarapuk">Apiradee Intarapuk</name><name sortKey="Pinyoowong, Danai" sort="Pinyoowong, Danai" uniqKey="Pinyoowong D" first="Danai" last="Pinyoowong">Danai Pinyoowong</name></noCountry><country name="Thaïlande"><noRegion><name sortKey="Pechgit, Prapassorn" sort="Pechgit, Prapassorn" uniqKey="Pechgit P" first="Prapassorn" last="Pechgit">Prapassorn Pechgit</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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