scholarly journals Laboratory Diagnosis of Dengue Virus Infection by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR) and IgM-Capture Enzyme-Linked Immunosorbent Assay (ELISA)

1999 ◽  
Vol 52 (4) ◽  
pp. 150-155
Author(s):  
Ken-Ichiro Yamada ◽  
Masaru Nawa ◽  
Tomohiko Takasaki ◽  
Sadao Yabe ◽  
Ichiro Kurane
Keyword(s):  
Polymerase Chain Reaction ◽  
Virus Infection ◽  
Dengue Virus ◽  
Reverse Transcriptase ◽  
Laboratory Diagnosis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Dengue Virus Infection ◽  
Polymerase Chain

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 Serological evidence of canine influenza virus infection in shelter dogs in Turkey

2018 ◽  
Vol 74 (6) ◽  
pp. 5986-2018 ◽  
Author(s):  
HAKAN AYDIN ◽  
AKIN KIRBAS ◽  
MEHMET OZKAN TIMURKAN ◽  
MUSTAFA SINAN AKTAS ◽  
GULIZAR ACAR KIRMIZI ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Influenza Virus ◽  
Virus Infection ◽  
Reverse Transcriptase ◽  
Influenza Virus Infection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Reaction ◽  
Canine Influenza Virus ◽  
Polymerase Chain ◽  
Canine Influenza

Influenza virus infection is an important disease which occurs in humans and a variety of animals. Because of the wide host adaptation and segmented genome, there is always the possibility of mutations and interspecies transmission of the influenza virus. Our study is the first to draw attention to canine influenza infection in Turkey. For this purpose, 208 sera and swab samples were collected from dogs with respiratory and nonrespiratory signs in various seasons. Out of the 208 dogs, 94 (45.2%) were male and 114 (54.8%) were female; the average age was 4.7 years. A total of 208 sera samples were tested for the presence of canine influenza virusspecific antibodies by the indirect enzyme-linked immunosorbent assay. The seroprevalence of canine influenza virus infection was 11/208 (5.8%). With regard to seasonal distribution, the highest rate of seropositivity was detected in spring, and the lowest in summer. Molecular detection of the canine influenza virus from nasal swab samples was done by reverse transcriptase polymerase chain reaction using specific universal primers for the hemagglutinin gene. Influenza virus nucleic acid could not be detected by reverse transcriptase polymerase chain reaction. In this study, we revealed for the first time the existence of the canine influenza virus in Turkey. Although the seroprevalence was relatively low, it would be useful to investigate the canine influenza virus on a large scale and among dogs with infectious respiratory disease in the Turkish dog population..

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 Virus Nipah

2016 ◽  
Vol 8 (2) ◽  
Author(s):  
Novie H. Rampengan
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Immunosorbent Assay ◽  
Nipah Virus ◽  
Specific Treatment ◽  
The Philippines ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

Abstract: Nipah virus caused outbreaks in Malaysia and Singapore in 2009 with a high mortality rates. It also erupted in Bangladesh, India, and the Philippines. Nipah virus infection varies from asymptomatic to severe manifestation with a mortality rate varies from 38% to 80%. Diagnosis can be confirmed by using reverse transcriptase-polymerase chain reaction (RT-PCR), immunohistochemical examination, enzyme-linked immunosorbent assay (ELISA), and neutralization test. There is still neither vaccine nor specific treatment for the Nipah virus so farKeywords: Nipah virus, signs and symptoms, diagnosisAbstrak: Virus Nipah menimbulkan outbreak di Malaysia dan Singapura tahun 2009 dengan angka kematian yang tinggi. Selain itu virus Nipah juga menimbulkan outbreak di Bangladesh, India, dan Filipina. Infeksi virus Nipah dapat bervariasi dari asimtomatik sampai bermanifestasi klinis yang berat dengan angka kematian bervariasi dari 38%-80%. Diagnosis dapat ditegakkan dengan reverse transcriptase-polymerase chain reaction (RT-PCR), pemeriksaan imunohistokimia, enzyme-linked immunosorbent assay (ELISA), dan tes netralisasi. Sampai saat ini belum ada vaksin dan terapi spesifik untuk virus Nipah.Kata kunci: virus Nipah, gejala, diagnosis

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 Evaluation of Sandwich Enzyme-Linked Immunosorbent Assay and Reverse Transcription Polymerase Chain Reaction for the Diagnosis of Foot-and-Mouth Disease

Intervirology ◽  
2021 ◽  
pp. 1-6
Author(s):  
Salman Khan ◽  
Syed Asad Ali Shah ◽  
Syed Muhammad Jamal
Keyword(s):  
Polymerase Chain Reaction ◽  
Viral Genome ◽  
Foot And Mouth Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Kappa Value ◽  
Polymerase Chain ◽  
Pcr Assays ◽  
Level Of Agreement

<b><i>Background:</i></b> Foot-and-mouth disease (FMD) is an infectious and highly contagious disease of cloven-hoofed domestic and wild animals, causing heavy economic losses to the livestock industry. Rapid and reliable diagnosis of the disease is essential for the implementation of effective control measures. This study compared sandwich enzyme-linked immunosorbent assay (S-ELISA) and conventional reverse transcription polymerase chain reaction (RT-PCR) for the diagnosis of FMD. <b><i>Methods:</i></b> A total of 60 epithelial samples from suspected cases of FMD were tested using both S-ELISA and RT-PCR assays. The level of agreement between the assays was assessed by calculating the Kappa value. <b><i>Results:</i></b> S-ELISA detected 38 (63%) samples positive for FMD virus (FMDV). Being predominant, serotype O was detected in 22 (57.9%) of the total samples tested positive, whereas 9 (23.7%) and 7 (18.4%) samples were found positive for serotypes A and Asia-1, respectively. RT-PCR detected viral genome in 51 (85%) of the samples using pan-FMDV primers set, 1F/1R. Thirty-six samples were found positive and 7 negative by both the tests. The level of agreement between the tests was assessed by calculating the Kappa value, which was found to be fair (Kappa value = 0.303 and 95% CI = 0.089; 0.517) and significant (<i>p</i> = 0.009). However, 2 samples, which were found positive on S-ELISA tested negative on RT-PCR. This may be attributed to the presence of nucleotide mismatch(es) in the primer-binding sites that may have resulted in failure of amplification of the viral genome. The serotype-specific RT-PCR assays not only confirmed serotyping results of S-ELISA but were also able to establish serotype in 9 S-ELISA-negative but pan-FMDV RT-PCR-positive samples. <b><i>Conclusions:</i></b> The RT-PCR assay contributes significantly to establishing a quick, sensitive, and definitive diagnosis of FMD in resource-constrained countries. Samples giving negative results in S-ELISA should be tested in RT-PCR for the disease detection and virus typing.

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