scholarly journals COMPARISON AND EVALUATION OF ONE-STEP REVERSE TRANSCRIPTASE-POLYMERASE CHAIN REACTION (RT-PCR) AND REVERSE TRANSCRIPTASE LOOP-MEDIATED ISOTHERMAL AMPLIFICATION ASSAY(RT-LAMP) FOR RAPID DETECTION OF FOOT-AND-MOUTH DISEASE VIRUS (FMDV)

2017 ◽  
Vol 1 (2) ◽  
pp. 16-19
Author(s):  
Waqas Ali ◽  
Mudasser Habib ◽  
Sanaullah Sajid ◽  
Rai Shafqat Ali Khan ◽  
Muhammad Ashir Zia ◽  
...  
1999 ◽  
Vol 80 (1) ◽  
pp. 45-52 ◽  
Author(s):  
Veluvarti Suryanarayana ◽  
Boynapalli Madanamohan ◽  
Pradeep Bist ◽  
Chittatoor Natarajan ◽  
Jon Duri Tratschin

2021 ◽  
Vol 22 (2) ◽  
pp. 189-197
Author(s):  
Putu Ayu Asri Damayanti ◽  
I Nyoman Mantik Astawa ◽  
Anak Agung Ayu Mirah Adi ◽  
I Made Sudarmaja ◽  
I Kadek Swastika ◽  
...  

Mosquito-specific viruses (MSVs) adalah virus yang hanya dapat bereplikasi pada sel nyamuk. Virus ini terdiri dari berbagai genus, salah satunya yang paling banyak ditemukan adalah dari famili Flaviviridae, genus Flavivirus. Namun, data keberadaan dan karakteristik MSVs dan vektornya di Bali saat ini sangat terbatas. Oleh karena itu, pengamatan untuk memperluas penemuan keragaman vektor dan filogenetik MSVs famili Flaviviridae, genus Flavivirus di Bali dilakukan pada tahun 2016-2018. Nyamuk-nyamuk dewasa ditangkap menggunakan light trap dan dikelompokkan berdasarkan spesies. Isolasi dan propagasi virus dilakukan pada galur sel C6/36 dan baby hamster kidney-21 (BHK-21). Identifikasi virus dilakukan dengan menggunakan one step reverse-transcriptase polymerase chain reaction (RT-PCR). Terdapat dua pool yang berasal dari nyamuk Anopheles vagus menampakan cythopathic effect (CPE) hanya pada galur sel C6/36 dari total 158 pool. Virus yang diisolasi memiliki persentase identity sekuen nukleotida tertinggi 97% dan sekuen asam amino 96% dengan virus Culex theileri Flavivirus isolat JKT-8650 yang diisolasi pada tahun 1981. Selanjutnya, virus dinamakan Mosquito Flavivirus Isolate Bali (MFB) dengan accession numbers KY995166 dan KY290258. Analisis filogenetik menunjukan bahwa MFB berada satu kluster dengan Culex theileri Flavivirus (CTFV) dari Indonesia, Culex Flavivuruses-Myanmar, Culex theileri Flavivirus-Portugal, dan Mosquito Flavivirus-Turki. Terdapat delapan nukelotida dan enam asam amino yang berbeda antara MFB dan CTFV Indonesia. Pada penelitian ini dapat disimpulkan bahwa MSVs dari famili Flaviviridae, genus Flavivirus berhasil diisolasi dari nyamuk An. vagus di Bali.


Author(s):  
A Kadir ◽  
S Ahmed

Foot and mouth disease (FMD) is a major constraint for livestock in Bangladesh; as outbreak of FMD remains uncontrolled despite vaccination. Accurate and Rapid detection of FMD virus with its serotype in field samples is indispensible. So, molecular detection of FMDV was adopted using RT-PCR (reverse transcription- polymerase chain reaction) and mRT-PCR (multiplex reverse transcription-polymerase chain reaction). Ten (10) FMD suspected clinical samples from cattle of two different outbreak areas of Sylhet district of Bangladesh was collected. One set of universal primer (P32:P33) was used in RT-PCR for the detection of FMD virus regardless of their serotypes and a cocktail of primer mix (P38:P40:P74-77:P110) was used in mRT-PCR intending the identification of the serotypes A, O, C and Asia 1. Using universal primer sets 90% of the samples generated amplicon of expected size, indicating the samples containing FMD virus. By mRT-PCR, two serotypes, ‘O’ and Asia 1 were successfully, whereas type C and A were absent in this study. Out of the 9 viruses; 7 was identified as serotype ‘O’ and 2 were identified as Asia-1. Our study indicates that FMDV serotype ‘O’ and Asia-1 was circulating in the two upazilas (Sub-district) of Sylhet district during the study period. Our study also endorses that, RT-PCR and mRT-PCR can successfully be used for a dependable and rapid detection of FMD. However, presence and detection of ‘O’ and Asia-1 serotype of FMDV through this study and serotype A by other researchers emphasizes the critical need for use of trivalent vaccine in the field.International Journal of Natural Sciences (2014), 4(1) 28-34


2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Fatemeh Khatami ◽  
Mohammad Saatchi ◽  
Seyed Saeed Tamehri Zadeh ◽  
Zahra Sadat Aghamir ◽  
Alireza Namazi Shabestari ◽  
...  

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.


Intervirology ◽  
2021 ◽  
pp. 1-6
Author(s):  
Salman Khan ◽  
Syed Asad Ali Shah ◽  
Syed Muhammad Jamal

<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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