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

2018 ◽  
Vol 64 (6) ◽  
pp. 393-400 ◽  
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.

Deteksi Transmisi Transovarial Virus Dengue PadaAedes AegyptiPada Wilayah Endemis Kota Pontianak

2021 ◽  
Vol 1 ◽  
pp. 984-995
Author(s):  
Malik Saepudin ◽  
Heru Subaris Kasjono ◽  
M Martini
Keyword(s):  
Polymerase Chain Reaction ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Aedes Albopictus ◽  
Transovarial Transmission ◽  
Chain Reaction ◽  
Cross Sectional ◽  
Transmission Index ◽  
Virus Serotype ◽  
Polymerase Chain

AbstractDengue virus is highly pathogenic in humans and spreads rapidly through Aedes aegypti and Aedes albopictus mosquitoes. More than half a billion people from 100 countries in the world are at serious risk of dengue virus infection. The purpose of this study was to prove the existence of transovarial transmission of dengue virus in Aedes aegypti mosquitoes with a transovarial transmission index (TTI) in endemic areas in Pontianak, West Kalimantan. This research method is descriptive observational with a cross-sectional study. The results of the Microscopic Examination of Head Squash Preparations on the Aedes aegypti mosquito, showed the Transovarial Transmission Index in Batulayang Village was 39.60% higher, compared to Sungai Jawi Village, which was 29.30%, but both were still lower than ITT in 2012. The results of the Aedes mosquito examination aegypti using the Polymerase Chain Reaction Transcription Reaction (PCR-TR) method found the dengue virus strain. The conclusion of this study proves that the transovarial transmission of dengue virus in Aedes aegypti mosquitoes in Sungai Jawi Village is 29.30% lower than in Batulayang Village by 39.60%, and the dengue virus serotype, DENV-3, has been found.Keywords: transovarial transmission; dengue virus; Aedes aegypti AbstrakVirus Dengue sangat patogen pada manusia dan menyebar dengan cepat melalui nyamuk Aedes aegypti dan Aedes albopictus. Lebih dari setengah miliar penduduk dari 100 negara di dunia berada pada risiko serius infeksi virus dengue. Tujuan dari penelitian ini adalah untuk membuktikan adanya penularan virus dengue transovarial pada nyamuk Aedes aegypti dengan indeks transmisi transovarial (TTI) di daerah Endemis di Pontianak, Kalimantan Barat. Metode penelitian ini adalah deskriptif observasional dengan studi potong lintang. Hasil Pemeriksaan Mikroskopis Sediaan Head Squash pada nyamuk Aedes aegypti, menunjukan Indeks Transmisi Transovarial pada Kelurahan Batulayang lebih tinggi yaitu 39,60%, dibandingkan dengan Kelurahan Sungai Jawi yaitu 29,30%, tetapi keduanya masih lebih rendah dibandingkan ITT pada Tahun 2012. Hasil pemeriksaan nyamuk Aedes aegypti dengan metode Polymerase Chain Reaction Transcription Reaction(PCR-TR) ditemukan strain virus Dengue-3. Kesimpulan penelitian ini membuktikan adanya transmisi transovarial virus dengue pada nyamuk Aedes aegypti di Kelurahan Sungai Jawi sebesar 29,30% lebih rendah dibandingkan di kelurahan Batulayang sebesar 39,60%, serta berhasil ditemukan serotipe virus Dengue yaitu DENV-3.Kata Kunci : transmisi transovarial; virus dengue; Aedes aegypti

scholarly journals REAL TIME POLYMERASE CHAIN REACTION (RT-PCR) FOR DNA PIG CONTAMINATION IDENTIFICATION SAUSAGE IN SIX DISTRICT PANDEGLANG BANTEN

2021 ◽  
Vol 1 (1) ◽  
pp. 81-88
Author(s):  
Hadi Susilo
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Meat Product ◽  
Meat Products ◽  
Pcr Analysis ◽  
Processed Meat ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Dna Purity ◽  
Polymerase Chain

Sausage is a meat product processed that is popular food especially in Pandeglang, Banten Province. The importance of halal certificates or the existence of the MUI (Indonesian Ulama Council) halal logo for processed meat products makes Muslim people confident to consume them. The aim this research was to identify pig DNA contamination in sausage products in six  districts in Pandeglang without the MUI halal labels using RT-PCR (Real Time-Polymerase Chain Reaction). RT PCR that can calculate to pig to fill these sample free from pig contamination. This research was divided into two stage, the first stage is extracted or carried out DNA and the second stage is RT PCR analysis. The results of the DNA purity test on sausage samples had DNA purity values ​​of 1.84-1.9 (A260 / A280) and resulted in sample concentrations ranging from 37.8 to 102.5 ng / µl.  The only amplification on the FAM curve was in the positive control pig.  the Cq value ranges from 30 - 31.29. The results of RT PCR on sausage samples in the district area in Pandeglang Banten did not detect the presence of pig DNA.

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

2018 ◽  
Vol 34 (2) ◽  
pp. 147-150 ◽  
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.

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

PLoS ONE ◽  
2021 ◽  
Vol 16 (9) ◽  
pp. e0257781
Author(s):  
Meng-Jia Lau ◽  
Ary A. Hoffmann ◽  
Nancy M. Endersby-Harshman
Keyword(s):  
Polymerase Chain Reaction ◽  
Aedes Aegypti ◽  
Relative Density ◽  
Real Time ◽  
Chain Reaction ◽  
Efficient Estimate ◽  
Polymerase Chain ◽  
Practical Implications

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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