Detection of dengue virus serotype 3 by reverse transcription-polymerase chain reaction in Aedes aegypti (Diptera, Culicidae) captured in Manaus, Amazonas

Abstract: Dengue hemorrhagic fever is an acute disease with clinical manifestations of hemorrhage caused by dengue virus infection. Manado is endemic dengue. Dengue virus has the ability to maintain its existence in nature through horizontal and vertical transmission. There are several ways to detect the dengue virus by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and immunohistochemistry Streptavidin Biotin Peroxidase Complex (ISBPC). This research aims to determine the wild Aedes aegypti population in Manado and to detect dengue virus in wild mosquito Aedes aegypti by ISBPC methods. This was a descriptive survey study with a cross sectional design to describe the transmission of dengue virus in wild mosquito Aedes aegypti in the city of Manado. The results showed that there were 5 wild Aedes aegypti mosquitoes positive for dengue virus, and 36 wild Aedes aegypti mosquitoes negative containing dengue virus. Conclusion: Of the 41 samples immunohistochemistry tested, 5 samples showed dengue virus transmission in wild mosquito Aedes aegypti in Manado which is a positive possibility of horizontal transmission.Keywords: detection of dengue virus, transmission, wild Aedes aegypti, Manado. Abstrak: Demam berdarah dengue adalah suatu penyakit akut dengan manifestasi klinis perdarahan yang disebabkan oleh infeksi virus dengue. Manado merupakan daerah endemis demam berdarah. Virus dengue memiliki kemampuan untuk mempertahankan keberadaannya di alam melalui transmisi horizontal dan vertikal. Ada beberapa cara untuk mendeteksi virus dengue yaitu Reverse Transcription-Polymerase Chain Reaction (RT-PCR) dan imunohistokimia Streptavidin Biotin Peroxidase Complex (SBPC). Penelitian ini bertujuan untuk mengetahui populasi nyamuk wild Aedes aegypti di Kota Manado dan mendeteksi virus dengue pada nyamuk wild Aedes aegypti dewasa menggunakan metode imunohistokimia streptavidin biotin peroxidase complex (ISBPC). Jenis penelitian ialah survei deskriptif dengan desain potong lintang untuk mengetahui gambaran transmisi virus dengue pada nyamuk wild Aedes aegypti betina di Kota Manado. Hasil pene;itian mendapatkan 5 nyamuk wild Aedes aegypti positif mengandung virus dengue, dan 36 nyamuk wild Aedes aegypti negatif mengandung virus dengue. Simpulan: Berdasarkan hasil penelitian dapat disimpulkan bahwa dari 41 sampel yang telah diuji imunohistokimia, 5 sampel gambaran transmisi virus dengue pada nyamuk wild Aedes aegypti betina di Kota Manado yang kemungkinan transmisi horizontal adalah positif. Kata kunci: deteksi virus dengue, transmisi, wild Aedes aegypti, Manado.

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Deteksi Transmisi Transovarial Virus Dengue PadaAedes AegyptiPada Wilayah Endemis Kota Pontianak

Prosiding Seminar Nasional Kesehatan ◽

10.48144/prosiding.v1i.779 ◽

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

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Use of immunodot blot and multiplex reverse transcriptase-polymerase chain reaction in dengue virus detection in macerates of Aedes aegypti larvae

Microbiology Research ◽

10.4081/mr.2012.e13 ◽

2012 ◽

Vol 3 (1) ◽

pp. 13

Author(s):

Aline T.A. Chagas ◽

Michelle D. Oliveira ◽

Jose M.S. Mezencio ◽

Eduardo A.M. Silva ◽

Leandro L. Oliveira ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Aedes Aegypti ◽

Dengue Virus ◽

Reverse Transcriptase ◽

Epidemiological Surveillance ◽

Rt Pcr ◽

Chain Reaction ◽

Denv Serotypes ◽

Polymerase Chain ◽

Pcr Techniques

The <em>Dengue virus</em> is the main arbovirus that affects man in terms of morbidity and mortality. The detection of the virus is very important for epidemiological surveillance, so here we propose to standardize and compare the immunodot blot (IDB) and multiplex reverse transcriptase-polymerase chain reaction (M-RT-PCR) techniques to detect and characterize the dengue virus (DENV) serotypes in samples of <em>Aedes aegypti</em> larvae. Thus, the IDB and M-RT-PCR techniques were standardized using macerated samples of larvae collected in nature. The use of monoclonal antibodies in IDB has not shown great results, but DENV detection through this method was possible using polyclonal antibodies. The distinction of serotypes 1, 2 and 3 was carried out by M-RT-PCR.

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Novel pan-serotype control RNA for dengue virus typing through real-time reverse transcription-polymerase chain reaction

Journal of Virological Methods ◽

10.1016/j.jviromet.2019.113677 ◽

2019 ◽

Vol 271 ◽

pp. 113677

Author(s):

Diego A. Álvarez-Díaz ◽

Paula A. Quintero ◽

Dioselina Peláez-Carvajal ◽

Nadim J. Ajami ◽

Jose A. Usme-Ciro

Keyword(s):

Polymerase Chain Reaction ◽

Dengue Virus ◽

Real Time ◽

Reverse Transcription ◽

Chain Reaction ◽

Polymerase Chain ◽

Virus Typing

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Circulation of Dengue Virus Serotypes in the City of Makkah, Saudi Arabia, as Determined by Reverse Transcription Polymerase Chain Reaction

Canadian Journal of Infectious Diseases and Medical Microbiology ◽

10.1155/2017/1646701 ◽

2017 ◽

Vol 2017 ◽

pp. 1-5 ◽

Cited By ~ 5

Author(s):

Sameer R. Organji ◽

Hussein H. Abulreesh ◽

Gamal E. H. Osman

Keyword(s):

Polymerase Chain Reaction ◽

Saudi Arabia ◽

Dengue Virus ◽

Dengue Fever ◽

Reverse Transcription ◽

Chain Reaction ◽

Denv Serotypes ◽

Polymerase Chain ◽

Low Prevalence ◽

The City

The present study was aimed to investigate the circulation of four dengue virus (DENV) serotypes in Makkah, Western Saudi Arabia. Blood samples were collected from 25 dengue fever-suspected patients and were subjected to molecular typing for DENV-1, DENV-2, DENV-3, and DENV-4 serotypes of dengue virus, by reverse transcription polymerase chain reaction (RT-PCR), using six sets of primers. Of the 25 samples, only six samples (24%) were found to be positive for dengue virus infection. The prevalence of DENV-1 was higher (50% of DENV-positive samples), as compared to DENV-2 (33.3%) and DENV-3 (16.6%) serotypes. The fourth serotype, DENV-4, was not detected in any of the DENV-positive samples. Although Makkah is considered endemic to dengue fever, we observed low prevalence of dengue virus in the city, which may be attributed to various factors. Nonetheless, the results presented herein confirm the circulation of DENV serotypes in the Western region of Saudi Arabia. To the best of our knowledge, the current study so far is the first report demonstrating the prevalence of the DENV-1 serotype in the city Makkah, Saudi Arabia.

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Deployable, Field-Sustainable, Reverse Transcription-Polymerase Chain Reaction Assays for Rapid Screening and Serotype Identification of Dengue Virus in Mosquitoes

Military Medicine ◽

10.7205/milmed.172.3.329 ◽

2007 ◽

Vol 172 (3) ◽

pp. 329-334 ◽

Cited By ~ 2

Author(s):

James C. McAvin ◽

Michael D. Powers ◽

Jamie A. Blow ◽

John L. Putnam ◽

William B. Huff ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Dengue Virus ◽

Reverse Transcription ◽

Rapid Screening ◽

Chain Reaction ◽

Polymerase Chain ◽

Serotype Identification

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Precipitation Solvents for RNA Extraction of Dengue Virus Type 3: Dimethylformamide, Ethylenediamintetraacetic Acid, and Ultrapure H2O

Jurnal Kimia Riset ◽

10.20473/jkr.v3i2.9353 ◽

2019 ◽

Vol 3 (2) ◽

pp. 78

Author(s):

Rizqidhana Juliana Putri ◽

Teguh Hari Sucipto ◽

Harsasi Setyawati ◽

Siti Churrotin ◽

Ilham Harlan Amarullah ◽

...

Keyword(s):

Polymerase Chain Reaction ◽

Dengue Virus ◽

Dengue Fever ◽

Reverse Transcription ◽

Rna Extraction ◽

Chain Reaction ◽

Dengue Virus Type 3 ◽

Pcr Method ◽

Polymerase Chain ◽

Type 3

Dengue is a disease caused by a virus from the family Flaviviradae, carried by a female mosquito of Aedes aegypti species. Dengue fever is widespread in the tropic areas. It caused by rainfall, temperature and unplanned urbanization. According to the ministry of health , almost all provinces in Indonesia are endemic areas of dengue fever. In 2014, up to mid-December Dengue Hemorrhagic Fever (DHF) patients in 34 provinces in Indonesia are 71,668 people and 641. This figure is lower than the previous year, 2013 with 112,511 people and 871 deaths . This disease consists of four types of serotypes, namely DENV-1, DENV-2, DENV-3, and DENV-4. This disease can be identified using a variety of methods, one of the method is Reverse Transcription - Polymerase Chain Reaction (RT-PCR) method. This study aims to determine the ability of Dimethylformamide (DMF), Ethylenediamintetraacetic Acid (EDTA), and Ultrapure H2O as the substitute of Ethanol for precipitation in RNA extraction process. The sample used in this research obtained from Surabaya. RNA extraction itself can be done by using a special kit for RNA extraction. In Reverse Transcription - Polymerase Chain Reaction method, first RNA is extracted and then transcribed back (Reverse Transcription) which then form cDNA that later will be amplified by using PCR method. In this study used specific primers for dengue virus type 3 (DENV-3). The results of this study show that DMF, EDTA, and Ultrapure H2O can be used as the substitute of Ethanol for precipitation on RNA extraction. The result is evidenced by the formation of viral DNA bands on gel electrophoresis results.

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Detection of Dengue Virus In Aedes sp. Mosquito at Home of DHF Patients in Ternate City

PROMOTIF Jurnal Kesehatan Masyarakat ◽

10.31934/promotif.v7i2.541 ◽

2017 ◽

Vol 7 (2) ◽

pp. 158-166

Author(s):

Mustafa Mustafa

Keyword(s):

Polymerase Chain Reaction ◽

Aedes Aegypti ◽

Dengue Virus ◽

Aedes Albopictus ◽

Chain Reaction ◽

Pcr Polymerase Chain Reaction ◽

Polymerase Chain ◽

At Home

Virus Dengue adalah penyebab penyakit demam berdarah dengue (DBD), ditularkan melalui gigitan nyamuk Aedes aegypti dan Aedes albopictus. Penyakit Demam Berdarah Dengue (DBD) adalah penyakit infeksi yang disebabkan oleh virus Dengue dengan serotipe DEN-1, DEN-2, DEN-3 dan DEN-4. Penelitian ini bertujuan untuk mengetahui keberadaan virus Dengue pada nyamuk Aedes sp di rumah penderita DBD Kota Ternate. Jenis penelitian yang digunakan adalah penelitian Deskriptif dengan uji laboratorium. Sampel pada penelitian ini adalah nyamuk jenis Aedes sp yang ditangkap dirumah penderita DBD. Teknik pengambilan sampel pada penelitian ini adalah Total sampling. Hasil penelitian ini menunjukkan kondisi lingkungan rumah penderita rata-rata berpotensial untuk perkembangan nyamuk dan jentik, suhu dalam rumah (91,67%), suhu luar rumah (83,33%), suhu air (91,67%), kelembaban (83,33%), pH air (75%) dan pencahayaan (91,67%). Jenis wadah berpotensial untuk perkebangan jentik yaitu bak mandi/WC 57,1%, drum plastik 40 % dan ember 36,8 %. Dari hasil pemeriksaan PCR (Polymerase Chain Reaction) pada 369 ekor nyamuk Aedes sp tidak menunjukkan adanya nyamuk yang mengandung virus Dengue-1. Kesimpulan dari penelitian ini adalah rata-rata Kondisi lingkungan fisik potensial untuk perkembangan larva dan nyamuk. Jenis wadah bak paling tinggi positif jentik, dan dari semua sampel nyamuk yang diperiksa tidak satupun yang mengandung virus Dengue serotipe DEN1.

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