Cross-Platform Evaluation of Commercial Real-Time SYBR Green RT-PCR Kits for Sensitive and Rapid Detection of European Bat<i>Lyssavirus</i>Type 1

Garam, Carine Peytavin de; Schereffer, J.L.; Marchal, Clotilde; Robardet, Emmanuelle; Cliquet, Florence

doi:10.1155/2015/839518

Cited by 24 publications

(24 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…2012; PICARD-MEYER, 2015). In this study, the kit I was faster, with better melting curves showing better performance and was used to evaluate the analytical sensitivity of Systems A and B.…”

Section: Resultsmentioning

confidence: 99%

Canine distemper virus detection by different methods of One-Step RT-qPCR

et al. 2016

View full text Add to dashboard Cite

Three commercial kits of One-Step RT-qPCR were evaluated for the molecular diagnosis of Canine Distemper Virus. Using the kit that showed better performance, two systems of Realtime RT-PCR (RT-qPCR) assays were tested and compared for analytical sensitivity to Canine Distemper RESUMO Três kits comerciais de One-Step RT-qPCR foram avaliados para o diagnóstico molecular do Vírus da Cinomose Canina.Utilizando o kit que apresentou melhor desempenho, dois sistemas de RT-PCR em tempo real (RT-qPCR) foram comparados quanto à sensibilidade analítica na detecção do RNA do Vírus da Cinomose Canina:One-Step RT-qPCR (Sistema A) e One-Step RT-qPCR seguido da NESTED-qPCR (Sistema B).Os limites de detecção dos dois sistemas foram determinados utilizando diluição

show abstract

Section: Resultsmentioning

confidence: 99%

Canine distemper virus detection by different methods of One-Step RT-qPCR

et al. 2016

View full text Add to dashboard Cite

show abstract

“…Since the clinical diagnosis of rabies is difficult and poorly reliable, especially if the history of a bite by a rabid animal is not present (15); in these cases, specialized diagnosis of rabies is required. The failure of routine diagnostic are not in molecular methods.…”

Section: Discussionmentioning

confidence: 99%

Development and Validation of Heminested RT-PCR and qRT-PCR Assays for Comprehensive Detection of Rabies Virus in the Suspected Rabid Brain and Saliva Samples

Amiri

Fadajan

Rasooli

et al. 2019

Arch Clin Infect Dis

View full text Add to dashboard Cite

Background:Rabies virus (RV) is one of the most dangerous zoonotic disease and major public health problems in most of the world, especially underdeveloped countries. Rabies is preventable by proper vaccination, even shortly after exposure. Today, it seems a fast, sensitive, and reliable rabies diagnostic method is required, which might reduce the financial burden of inappropriate diagnosis. Objectives: The aim of this study was to develop and validate two molecular techniques, including heminested RT-PCR and qRT-PCR assays, for comprehensive detection of rabies virus in the suspected rabid brain and saliva samples. Methods: In this study, we developed qRT-PCR as a fast, sensitive, and specific method for rapid detection of rabies virus in brain and saliva samples. Also, the sensitivity and specificity of the method were compared with heminested RT-PCR test and direct fluorescent antibody (dFA) as a serologic gold standard method of World Health Organization (WHO) and MIT (mouse inoculation test) as a confirmatory test. Results: A combination of compatible primers based on RNA-dependent RNA polymerase (L) gene of the Pasteur virus fixed strain (PV) (accession number. M13215) was used for developing the qRT-PCR assay. Primer and probes were designed according to other Iran circulating viruses genomes that were available in public databases (GenBank). The clinical sensitivities of qRT-PCR and heminested RT-PCR methods were determined 97.14% and 94.3%, respectively. In addition, the clinical specificities of qRT-PCR and heminested RT-PCR methods were determined 93.75% and 88.24%, respectively. Also, the analytical sensitivities of qRT-PCR and heminested RT-PCR methods were about 5 × 10 2 and 5 × 10 3 FFU/mL, respectively. Conclusions:In this study, qRT-PCR assay as a diagnostic molecular method with high sensitivity and specificity was developed for the detection of the rabies virus genome in both brain and saliva samples. Therefore, this rapid, accurate, and cost-effective detection and quantification method may be used as an investigative tool, which can be valid for detection of target viral genome in the research and diagnosis field.

show abstract

“…For that study, HPV was selected to generate HPV RNA transcript. We selected two different commercial cDNA synthesis kits for detecting HPV and evaluated the results using efficiency and R 2 as shown in a paper [28]. Results showed that efficiency (%) was between 90%∼110%…”

Section: Discussionmentioning

confidence: 99%

“…Real-time PCR for HPV screening test is performed after RNA extraction, complementary DNA (cDNA) synthesis, and amplification of cDNA [25][26][27]. Real-time PCR gene expression can depend on efficiency (%) of complementary DNA synthesis even if total RNA is extracted efficiently [28].…”

Section: Introductionmentioning

confidence: 99%

Evaluation of Commercial Complementary DNA Synthesis Kits for Detecting Human Papillomavirus

Park

Chang

et al. 2019

Korean J Clin Lab Sci.

View full text Add to dashboard Cite

Cervical cancer is the fourth most common malignant neoplasm in women worldwide. Most cases of cervical cancer are caused by an infection by the human papillomavirus. Molecular diagnostic methods have emerged to detect the HPV for sensitivity, specificity, and objectivity. In particular, real-time PCR has been introduced to acquire a more sensitive target DNA or RNA. RNA extraction and complementary DNA synthesis are proceeded before performing real-time PCR targeting RNA. To identify an adequate and sensitive cDNA synthesis kit, this study evaluated the two commonly used kits for cDNA synthesis. The results show that the R 2 and efficiency (%) of the two cDNA synthesis kits were similar in the cervical cancer cell lines. On the other hand, the Takara kit compared to Invitrogen kit showed P＜0.001 in the 10 2 and 10 3 SiHa cell count. The Takara kit compared to the Invitrogen kit showed P＜0.001 in the 10 1 and 10 2 HeLa cell count. Furthermore, 8, 4, 2, 1, and 0.5 ml of forty exfoliated cell samples were used to compare the cDNA synthesis kits. The Takara kit compared to the Invitrogen kit showed P＜0.01 in 8, 4, and 1 ml and P＜0.05 in 0.5 mL. The study was performed to identify the most appropriate cDNA synthesis kit and suggests that a cDNA synthesis kit could affect the real-time PCR results.

show abstract

Cross-Platform Evaluation of Commercial Real-Time SYBR Green RT-PCR Kits for Sensitive and Rapid Detection of European BatLyssavirusType 1

Cited by 24 publications

References 23 publications

Canine distemper virus detection by different methods of One-Step RT-qPCR

Canine distemper virus detection by different methods of One-Step RT-qPCR

Development and Validation of Heminested RT-PCR and qRT-PCR Assays for Comprehensive Detection of Rabies Virus in the Suspected Rabid Brain and Saliva Samples

Evaluation of Commercial Complementary DNA Synthesis Kits for Detecting Human Papillomavirus

Contact Info

Product

Resources

About