scholarly journals Study on the Dynamic Proliferation of JEV in BHK-21 Cells

Intervirology ◽  
2021 ◽  
pp. 1-7
Author(s):  
Fuliang Zhang ◽  
Jun Luo ◽  
Man Teng ◽  
Guangxu Xing ◽  
Junqing Guo ◽  
...  
Keyword(s):  
Cell Suspension ◽  
Real Time ◽  
Japanese Encephalitis ◽  
Zoonotic Diseases ◽  
Virus Inoculation ◽  
Central Nervous System Damage ◽  
Virus Rna ◽  
Pcr Method ◽  
Culture Supernate ◽  
Culturing Conditions

<b><i>Introduction:</i></b> Epidemic Japanese encephalitis is one of the most important zoonotic diseases that cause central nervous system damage. The vaccination has become the most effective and economical measure for its control. Hence, real-time monitoring of Japanese encephalitis virus (JEV) proliferation is crucial to optimize virus inoculation, culturing conditions, and virus harvest time. <b><i>Methods:</i></b> The proliferation dynamics of JEV in BHK-21 cells was studied by combining the established quantitative PCR method with the conventional TCID<sub>50</sub> assay in this study. <b><i>Results:</i></b> The proliferation curve determined by the 2 methods has a definite parallel relationship, but the quantitative real-time PCR method (4 h) is faster and more sensitive than the TCID<sub>50</sub> method (3–4 days). The determination results of TCID<sub>50</sub> showed that the highest viral titer was 10<sup>5.44</sup> TCID<sub>50</sub>/0.1 mL and 10<sup>4.86</sup> TCID<sub>50</sub>/0.1 mL in cell suspension and culture supernate, respectively, while the virus RNA copies reached the peak at 1.0 × 10<sup>7.5</sup> copies/µL and 1.0 × 10<sup>5.6</sup> copies/µL in cell suspension and culture supernate, respectively. <b><i>Conclusion:</i></b> The comprehensive analysis showed that the best time for JEV proliferation in BHK-21 cell was 60 h post infection.

scholarly journals Absolute Quantitation of Different Genotypes of Hepatitis C Virus RNA in Clinical Samples by a Modified Real-Time PCR Method

2011 ◽  
Vol 42 (6) ◽  
pp. 333-337
Author(s):  
Karina del Carmen Trujillo-Murillo ◽  
Diana Cristina Pérez-Ibave ◽  
Clara Patricia Ríos-Ibarra ◽  
Eda Guadalupe Ramirez-Valles ◽  
Ana Rosa Rincón-Sánchez ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Real Time ◽  
Real Time Pcr ◽  
Clinical Samples ◽  
Hepatitis C Virus Rna ◽  
Absolute Quantitation ◽  
Virus Rna ◽  
Pcr Method

scholarly journals Hepatitis C Virus RNA Real-Time Quantitative RT-PCR Method Based on a New Primer Design Strategy

2016 ◽  
Vol 18 (1) ◽  
pp. 84-91 ◽  
Author(s):  
Lida Chen ◽  
Wenli Li ◽  
Kuo Zhang ◽  
Rui Zhang ◽  
Tian Lu ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Real Time ◽  
Design Strategy ◽  
Primer Design ◽  
Rt Pcr ◽  
Hepatitis C Virus Rna ◽  
Virus Rna ◽  
Pcr Method

A one step real time PCR method for the quantification of hepatitis delta virus RNA using an external armored RNA standard and intrinsic internal control

2014 ◽  
Vol 60 (1) ◽  
pp. 11-15 ◽  
Author(s):  
Ersin Karataylı ◽  
Yasemin Çelik Altunoğlu ◽  
Senem Ceren Karataylı ◽  
S. Gökçe K. Alagöz ◽  
Kubilay Çınar ◽  
...  
Keyword(s):  
Real Time ◽  
Internal Control ◽  
Real Time Pcr ◽  
Hepatitis Delta Virus ◽  
Hepatitis Delta ◽  
Virus Rna ◽  
One Step ◽  
Pcr Method ◽  
Armored Rna ◽  
Delta Virus

Detection of Streptococcus pneumoniae in Sputum Samples by Real- Time PCR.

2020 ◽  
Vol 18 ◽  
Author(s):  
Pegah Shakib ◽  
Mohammad Reza Zolfaghari
Keyword(s):  
Streptococcus Pneumoniae ◽  
Real Time ◽  
Real Time Pcr ◽  
False Negative ◽  
Cross Sectional Study ◽  
Laboratory Culture ◽  
Cross Sectional ◽  
Negative Results ◽  
False Negative Results ◽  
Pcr Method

Background: Conventional laboratory culture-based methods for diagnosis of Streptococcus pneumoniae are time-consuming and yield false negative results. Molecular methods including real-time (RT)-PCR rapid methods and conventional PCR due to higher sensitivity and accuracy have been replaced instead traditional culture assay. The aim of the current study was to evaluate lytA gene for detection of Streptococcus pneumoniae in the cerebrospinal fluid of human patients with meningitis using real-time PCR assay. Material and Methods: In this cross-sectional study, a total of 30 clinical specimens were collected from patients in a period from September to December 2018. In order to evaluate the presence of lytA gene, conventional and real-time PCR methods were used without culture. Results: From 30 sputum samples five (16.66%) isolates were identified as S. pneumoniae by lytA PCR and sequencing. Discussion: In this research, an accurate and rapid real-time PCR method was used, which is based on lytA gene for diagnosis of bacteria so that it can be diagnosed. Based on the sequencing results, the sensitivity for detection of lytA gene was 100% (5/5).

scholarly journals Improved real-time RT-PCR method for high-throughput measurements using second derivative calculation and double correction

BioTechniques ◽  
2005 ◽  
Vol 38 (2) ◽  
pp. 287-293 ◽  
Author(s):  
Van Luu-The ◽  
Nathalie Paquet ◽  
Ezequiel Calvo ◽  
Jean Cumps
Keyword(s):  
Real Time ◽  
High Throughput ◽  
Second Derivative ◽  
Rt Pcr ◽  
Pcr Method

scholarly journals Trueness and precision of the real-time RT-PCR method for quantifying the chronic bee paralysis virus genome in bee homogenates evaluated by a comparative inter-laboratory study

2017 ◽  
Vol 248 ◽  
pp. 217-225 ◽  
Author(s):  
Frank Schurr ◽  
Nicolas Cougoule ◽  
Marie-Pierre Rivière ◽  
Magali Ribière-Chabert ◽  
Hamid Achour ◽  
...  
Keyword(s):  
Real Time ◽  
Laboratory Study ◽  
Virus Genome ◽  
Rt Pcr ◽  
The Real ◽  
Pcr Method ◽  
Paralysis Virus

scholarly journals Real-Time PCR Assay for Detection and Enumeration of Dekkera bruxellensis in Wine

2003 ◽  
Vol 69 (12) ◽  
pp. 7430-7434 ◽  
Author(s):  
Trevor G. Phister ◽  
David A. Mills
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Pcr Primers ◽  
Rrna Gene ◽  
Dekkera Bruxellensis ◽  
Pcr Assay ◽  
Specific Pcr ◽  
Spoilage Yeast ◽  
Pcr Method ◽  
26S Rrna

ABSTRACT Traditional methods to detect the spoilage yeast Dekkera bruxellensis from wine involve lengthy enrichments. To overcome this difficulty, we developed a quantitative real-time PCR method to directly detect and enumerate D. bruxellensis in wine. Specific PCR primers to D. bruxellensis were designed to the 26S rRNA gene, and nontarget yeast and bacteria common to the winery environment were not amplified. The assay was linear over a range of cell concentrations (6 log units) and could detect as little as 1 cell per ml in wine. The addition of large amounts of nontarget yeasts did not impact the efficiency of the assay. This method will be helpful to identify possible routes of D. bruxellensis infection in winery environments. Moreover, the time involved in performing the assay (3 h) should enable winemakers to more quickly make wine processing decisions in order to reduce the threat of spoilage by D. bruxellensis.

Development and application of a novel reverse transcription real-time PCR method for miR-499 quantification

2013 ◽  
Vol 46 (15) ◽  
pp. 1566-1571 ◽  
Author(s):  
Weidong Zheng ◽  
Yuwei Di ◽  
Yinghong Liu ◽  
Ge Huang ◽  
Youwei Zheng ◽  
...  
Keyword(s):  
Real Time ◽  
Reverse Transcription ◽  
Real Time Pcr ◽  
Pcr Method
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