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

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.

scholarly journals Optimization of Real-time Reverse Transcriptase Polymerase Chain Reaction for Detection of Dengue Virus

2020 ◽  
pp. 1-7
Author(s):  
Kaunara A. Azizi ◽  
Arnold J. Ndaro ◽  
Athanasia Maro ◽  
Adonira Saro ◽  
Reginald A. Kavishe
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleic Acid ◽  
Dengue Virus ◽  
Reverse Transcriptase ◽  
Real Time ◽  
Acid Extraction ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Reliable Technique ◽  
Polymerase Chain

Aims: This study was set to optimize conditions for real time reverse transcriptase polymerase chain reaction (RT-PCR) for detection of dengue virus by using rapid and simple nucleic acid extraction method. Methodology: One step and two step real time RT-PCR were evaluated in different PCR thermocyclers. Extraction of viral RNA was done by using a simple boom method. Results: The real time RT-PCR technique was successfully optimized using simple and rapid method for purification of nucleic acid, ‘boom method’. The technique works better when performed in a two-step procedure and can works well with all range of real time PCR machines. The optimized real time RT-PCR used in the present study is a valuable and reliable technique for routine diagnosis of dengue. Further investigation on the cost effectiveness in adopting this technique for routine screening and monitoring of the dengue infection should be done.

scholarly journals IDENTIFICATION OF DENGUE VIRUS SEROTYPES AT THE DR. SOETOMO HOSPITAL SURABAYA IN 2016 AND ITS CORRELATION WITH NS1 ANTIGEN DETECTION

2018 ◽  
Vol 23 (2) ◽  
pp. 151
Author(s):  
Jeine Stela Akualing ◽  
Aryati Aryati ◽  
Puspa Wardhani ◽  
Usman Hadi
Keyword(s):  
Polymerase Chain Reaction ◽  
Dengue Virus ◽  
Reverse Transcriptase ◽  
Ribonucleic Acid ◽  
Rna Virus ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Ns1 Antigen ◽  
Dengue Virus Serotypes ◽  
Polymerase Chain

Serotipe virus dengue yang beredar terus mengalami perubahan dan berbeda di setiap daerah. Pergeseran serotipe maupun genotipedi dalamnya, mempengaruhi terjadinya wabah dengue di berbagai negara. Perbedaan serotipe diduga bernasab dengan deteksi antigen(Ag) non-structural 1 (NS1), namun belum banyak penelitian yang mendukung hal tersebut. Penelitian potong lintang dikerjakan sejakFebruari-Agustus 2016 dan didapatkan 60 subjek infeksi virus dengue (IVD) dan 25 non-IVD. Ribonucleic acid (RNA) virus denguediperiksa di semua subjek menggunakan Simplexa Dengue Real-Time Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR)termasuk identifikasi serotipe virus dengue dan pemeriksaan NS1 menggunakan uji cepat NS1 Panbio. Perbedaan perbandingan variabelkategorikal dianalisis dengan uji Fisher Exact. Kenasaban antara serotipe dengan deteksi Ag NS1 dianalisis dengan Chi-Kuadrat. RNAvirus dengue terdeteksi di 43 dari 60 subjek IVD (71,7%). Serotipe terbanyak adalah DENV-3 (62,8%). Pergeseran dominasi serotipetelah terjadi di Surabaya, sebelumnya dari DENV-2 ke DENV-1 dan sekarang DENV-3, kemungkinan akibat mobilitas pejamu, transporvirus dan faktor geografis. Kepekaan uji cepat NS1 75% dan kekhasan 100%. Persentase deteksi NS1 antar serotipe berbeda bermakna(p=0,002). Deteksi NS1 lebih rendah pada DENV-1 dibandingkan DENV-2 (p=0,007) ataupun DENV-3 (p=0,003). Serotipe virusdengue bernasab dengan deteksi NS1 (p=0,005). Ciri serotipe maupun genotipe virus dengue kemungkinan mempengaruhi sekresiNS1. Telah terjadi pergeseran serotipe virus dengue di pasien IVD di Surabaya sehingga diperlukan surveillance berkesinambunganuntuk memperkirakan terjadinya wabah. Serotipe bernasab dengan deteksi NS1. Salah satu penyebab hasil negatif palsu NS1 adalahperbedaan serotipe.

scholarly journals Deteksi transmisi virus dengue pada nyamuk wild Aedes Aegypti betina di Kota Manado

2016 ◽  
Vol 4 (2) ◽  
Author(s):  
Grace Trovancia ◽  
Angle Sorisi ◽  
Josef S.B. Tuda
Keyword(s):  
Polymerase Chain Reaction ◽  
Aedes Aegypti ◽  
Dengue Virus ◽  
Reverse Transcription ◽  
Clinical Manifestations ◽  
Virus Transmission ◽  
Survey Study ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

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.

scholarly journals Optimization Of Real-Time Reverse Transcriptase Polymerase Chain Reaction For Detection Of Dengue Virus

2020 ◽  
Author(s):  
Kaunara Ally Azizi ◽  
Arnold J Ndaro ◽  
Athanasia Maro ◽  
Adonira T Saro ◽  
Reginald Kavishe
Keyword(s):  
Polymerase Chain Reaction ◽  
Nucleic Acid ◽  
Dengue Virus ◽  
Reverse Transcriptase ◽  
Real Time ◽  
Diagnostic Methods ◽  
Virus Infections ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract Objective Rapid and accurate laboratory confirmatory is very essential for control measures of dengue virus infections. However, many cases of dengue virus infections in most of the hospitals remain undiagnosed due to presence of other febrile illnesses with overlapping symptoms and lack of specificity in most of laboratory diagnostic methods. This study was set to optimize conditions for real time reverse transcriptase polymerase chain reaction (RT-PCR) for detection of dengue virus by using rapid and simple nucleic acid extraction method.Results The real time RT-PCR technique was successfully optimized using simple and rapid method for purification of nucleic acid, ‘boom method’. The technique works better when performed in a two-step procedure and can works well with all range of real time PCR machines. The optimized real time RT-PCR used in the present study is a valuable and reliable technique for routine diagnosis of dengue. Further investigation on the cost effectiveness in adopting this technique for routine screening and monitoring of the dengue infection should be done.

scholarly journals A meta-analysis of accuracy and sensitivity of chest CT and RT-PCR in COVID-19 diagnosis

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Fatemeh Khatami ◽  
Mohammad Saatchi ◽  
Seyed Saeed Tamehri Zadeh ◽  
Zahra Sadat Aghamir ◽  
Alireza Namazi Shabestari ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Ct Scan ◽  
Predictive Value ◽  
Meta Analysis ◽  
Chest Ct ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Chest Ct Scan

AbstractNowadays there is an ongoing acute respiratory outbreak caused by the novel highly contagious coronavirus (COVID-19). The diagnostic protocol is based on quantitative reverse-transcription polymerase chain reaction (RT-PCR) and chests CT scan, with uncertain accuracy. This meta-analysis study determines the diagnostic value of an initial chest CT scan in patients with COVID-19 infection in comparison with RT-PCR. Three main databases; PubMed (MEDLINE), Scopus, and EMBASE were systematically searched for all published literature from January 1st, 2019, to the 21st May 2020 with the keywords "COVID19 virus", "2019 novel coronavirus", "Wuhan coronavirus", "2019-nCoV", "X-Ray Computed Tomography", "Polymerase Chain Reaction", "Reverse Transcriptase PCR", and "PCR Reverse Transcriptase". All relevant case-series, cross-sectional, and cohort studies were selected. Data extraction and analysis were performed using STATA v.14.0SE (College Station, TX, USA) and RevMan 5. Among 1022 articles, 60 studies were eligible for totalizing 5744 patients. The overall sensitivity, specificity, positive predictive value, and negative predictive value of chest CT scan compared to RT-PCR were 87% (95% CI 85–90%), 46% (95% CI 29–63%), 69% (95% CI 56–72%), and 89% (95% CI 82–96%), respectively. It is important to rely on the repeated RT-PCR three times to give 99% accuracy, especially in negative samples. Regarding the overall diagnostic sensitivity of 87% for chest CT, the RT-PCR testing is essential and should be repeated to escape misdiagnosis.

scholarly journals Deteksi Molekuler Virus Dengue dan Chikungunya pada Nyamuk Aedes spp. di Kecamatan Cilongok

2020 ◽  
Vol 2 (2) ◽  
pp. 181
Author(s):  
Dwi Iva Fitriana ◽  
Endang Srimurni Kusmintarsih ◽  
Trisnowati Budi Ambarningrum
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

DBD dan chikungunya merupakan salah satu penyakit yang masih menjadi masalah di Indonesia. Kecamatan Cilongok merupakan salah kecamatan endemis DBD dan pernah mengalami KLB chikungunya. Deteksi virus pada nyamuk sebelum menginfeksi manusia penting sebagai peringatan dini dalam upaya pencegahan wabah di daerah endemis. Tujuan penelitian ini adalah mengetahui infeksi virus Dengue dan Chikungunya pada nyamuk Aedes spp. yang ditangkap. Penelitian ini dilakukan di empat desa di Kecamatan Cilongok yang meliputi Desa Cilongok, Pernasidi, Kalisari, dan Jatisaba, pengambilan sampel dilakukan secara purposive. Deteksi virus Dengue dan Chikungunya pada nyamuk dilakukan menggunakan metode Reverse-Transcriptase Polymerase Chain Reaction (RT-PCR). Hasil positifitas virus dianalisis secara deskriptif untuk menggambarkan potensi transmisi virus. Hasil penelitian menunjukkan bahwa nyamuk Aedes spp. yang tertangkap tidak mengandung virus Dengue dan Chikungunya.  

scholarly journals A reverse transcriptase-polymerase chain reaction detects heterogeneous chimeric mRNAs in leukemias with 11q23 abnormalities

Blood ◽  
1994 ◽  
Vol 83 (10) ◽  
pp. 2912-2921 ◽  
Author(s):  
K Yamamoto ◽  
M Seto ◽  
S Iida ◽  
H Komatsu ◽  
N Kamada ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Response To Therapy ◽  
Chromosome 9 ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Single Clone ◽  
Sensitivity Studies ◽  
Polymerase Chain ◽  
11Q23 Abnormalities

Abstract The MLL gene involved in 11q23 translocations found in the majority of infantile leukemias and some secondary leukemias makes fusion transcripts with genes such as LTG4 (chromosome 4), LTG9 (chromosome 9), and LTG19 (chromosome 19) as a result of reciprocal translocation. We have examined 25 cases of leukemias with 11q23 abnormalities by Southern blot analysis and the reverse transcriptase-polymerase chain reaction (RT-PCR). Using various primer pairs, chimeric mRNAs could be amplified in 6 of 7 leukemias with t(4;11), 6 of 8 leukemias with t(9;11) including secondary leukemia, 8 of 9 leukemias with t(11;19), and 1 with a deletion at 11q23. The chimeric mRNAs were heterogeneous and differential usage of the MLL exons was found, irrespective of the partner chromosomes. Sensitivity studies showed that a single clone with chimeric mRNA in 10(4) to 10(5) cells could be detected. These findings show that the present RT-PCR settings provide a rapid, accurate, and sensitive tool for diagnosing leukemias with 11q23 translocations and for monitoring response to therapy in these patients.

Sign in / Sign up

Export Citation Format

Share Document