A diagnostic primer pair to distinguish between wMel and wAlbB Wolbachia infections

Detection of the Wolbachia endosymbiont in Aedes aegypti mosquitoes through real-time polymerase chain reaction assays is widely used during and after Wolbachia releases in dengue reduction trials involving the wMel and wAlbB strains. Although several different primer pairs have been applied in current successful Wolbachia releases, they cannot be used in a single assay to distinguish between these strains. Here, we developed a new diagnostic primer pair, wMwA, which can detect the wMel or wAlbB infection in the same assay. We also tested current Wolbachia primers and show that there is variation in their performance when they are used to assess the relative density of Wolbachia. The new wMwA primers provide an accurate and efficient estimate of the presence and density of both Wolbachia infections, with practical implications for Wolbachia estimates in field collected Ae. aegypti where Wolbachia releases have taken place.

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Report of Chikungunya Virus in Wild Populations of Aedes aegypti in Guerrero State, Mexico

Journal of the American Mosquito Control Association ◽

10.2987/17-6683.1 ◽

2018 ◽

Vol 34 (2) ◽

pp. 147-150 ◽

Cited By ~ 1

Author(s):

Gustavo Ponce-García ◽

Adriana E. Flores-Suarez ◽

Karina Villanueva-Segura ◽

Martha Lopez-Rodriguez ◽

Felipe Dzul ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Aedes Aegypti ◽

Reverse Transcriptase ◽

Real Time ◽

Chikungunya Virus ◽

Wild Populations ◽

Chain Reaction ◽

Male And Female ◽

San Marcos ◽

Polymerase Chain

ABSTRACT We detected vertical transmission of chikungunya virus (CHIKV) in wild populations of Aedes aegypti from San Marcos, Guerrero, Mexico, with real-time reverse transcriptase–polymerase chain reaction. A total of 20 pools (1–11 specimens/pool) of larvae, male, and female mosquitoes were tested. We report the detection of CHIKV in 2 of 11 larval pools, 4 of 5 male pools, and 1 of 4 female pools, from field-collected mosquitoes.

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ANALYSIS OF THE REPLICATION KINETICS OF THE CHIMERIVAX™-DEN 1, 2, 3, 4 TETRAVALENT VIRUS MIXTURE IN AEDES AEGYPTI BY REAL-TIME REVERSE TRANSCRIPTASE–POLYMERASE CHAIN REACTION

American Journal of Tropical Medicine and Hygiene ◽

10.4269/ajtmh.2004.70.89 ◽

2004 ◽

Vol 70 (1) ◽

pp. 89-97 ◽

Cited By ~ 36

Author(s):

BARBARA W. JOHNSON ◽

MARY B. CRABTREE ◽

TRUDY V. CHAMBERS ◽

BARRY R. MILLER ◽

THOMAS P. MONATH ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Aedes Aegypti ◽

Reverse Transcriptase ◽

Real Time ◽

Chain Reaction ◽

Polymerase Chain ◽

Kinetics Of

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Transovarial transmission of dengue 1 virus in Aedes aegypti larvae: real-time PCR analysis in a Brazilian city with high mosquito population density

Canadian Journal of Microbiology ◽

10.1139/cjm-2017-0614 ◽

2018 ◽

Vol 64 (6) ◽

pp. 393-400 ◽

Cited By ~ 1

Author(s):

Alexsander Moraes ◽

Filipe C. Cortelli ◽

Taís B. Miranda ◽

Davi R. Aquino ◽

José R. Cortelli ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Aedes Aegypti ◽

Real Time ◽

Pcr Analysis ◽

Transovarial Transmission ◽

Common Disease ◽

Chain Reaction ◽

Primary Mechanism ◽

Virus Serotype ◽

Polymerase Chain

Transovarial transmission is among the reported factors able to influence environmental maintenance of dengue virus (DENV). Endemic areas with active transmission of dengue are suitable for studying transovarial transmission. Brazil is a country where dengue is endemic and where DENV-1 is the most common disease-related virus serotype. This study aimed to identify transovarial transmission of DENV-1 in Aedes aegypti larvae by reverse-transcriptase nested real-time polymerase chain reaction. Between March and October 2016, Culicidae larvae were collected using traps in 3 locations in Taubaté, São Paulo, Brazil, which has a high occurrence of dengue. The collected larvae were sacrificed in the 3rd or 4th larval stage, classified, and stored at –20 °C. The A. aegypti larvae samples (n = 910) were separated into 91 pools of 10 specimens each from which RNA was extracted, reverse transcribed into cDNA, and analyzed by nested qPCR. None of the pools tested positive for DENV-1. Due to the absence of detectable virus in the evaluated samples, we concluded that transovarial transmission may not be the primary mechanism for maintenance of DENV-1 in this particular environment.

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