scholarly journals ECC–RT-PCR: a new method to determine the viability and infectivity of Giardia cysts

2012 ◽  
Vol 16 (5) ◽  
pp. e350-e353 ◽  
Author(s):  
Absar Alum ◽  
Basel Sbai ◽  
Hamas Asaad ◽  
Joseph R. Rubino ◽  
M. Khalid Ijaz
Keyword(s):  
New Method ◽  
Rt Pcr ◽  
Giardia Cysts

Magnetic bead RT-PCR: establishment of a new method for detecting circulating melanoma cells

2002 ◽  
Vol 12 (4) ◽  
pp. 309-317 ◽  
Author(s):  
J. Georgieva ◽  
A. Milling ◽  
C. E. Orfanos ◽  
C. C. Geilen
Keyword(s):  
Magnetic Bead ◽  
Melanoma Cells ◽  
New Method ◽  
Rt Pcr

Improvements in the RT-PCR detection of enteric viruses in environmental samples

1998 ◽  
Vol 38 (12) ◽  
pp. 83-86 ◽  
Author(s):  
K. J. Schwab ◽  
F. H. Neill ◽  
M. K. Estes ◽  
R. L. Atmar
Keyword(s):  
Nucleic Acid ◽  
Reverse Transcriptase ◽  
Environmental Samples ◽  
Internal Standard ◽  
Pcr Amplification ◽  
Enteric Viruses ◽  
New Method ◽  
Rt Pcr ◽  
Pcr Product ◽  
Enzyme Method

Current methods for the detection of nucleic acid from enteric viruses in environmental samples usually involve extensive concentration and purification of target viruses followed by RT-PCR amplification using two enzymes, reverse transcriptase and Taq polymerase. We have developed a modified method that improves RT-PCR assays by: (i) the use of an RT-PCR internal standard control RNA to identify potential false negative results caused by inhibition of RT-PCR enzymes; (ii) the use of rTth (Perkin-Elmer, Foster City, CA), a heat-stable enzyme that functions as both a reverse transcriptase and DNA polymerase in a single-tube, single-buffer, elevated-temperature reaction; and (iii) the use of thermolabile uracil N-glycosylase (HK-UNG) (Epicentre Technologies, Madison, WI) to prevent PCR product carryover contamination. The new method was compared to the traditional two-enzyme, RT-PCR method for detection of Norwalk virus (NV) and hepatitis A virus (HAV) in buffer, stool, clam and oyster samples. The new method was at least as sensitive in NV and HAV detection compared to the traditional two-enzyme method. The internal standard control successfully detected inhibitors to RT-PCR amplification. NV and HAV PCR products generated with dUTP replacing dTTP during amplification were seeded into subsequent samples to test the prevention of PCR product carryover contamination by HK-UNG. The new method successfully eliminated PCR product carryover contamination in contrast to the traditional two-enzyme method. These improvements to viral nucleic acid detection have the potential to improve sensitivity, specificity and confidence in RT-PCR results.

Real time RT-PCR quantification: a new method to study hepatitis C virus adsorption on vero cells

2001 ◽  
Vol 34 ◽  
pp. 117
Author(s):  
R. Germi ◽  
J.M. Crance ◽  
D. Garin ◽  
J. Guimet ◽  
C. Rothlisberger ◽  
...  
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Real Time ◽  
Vero Cells ◽  
New Method ◽  
Rt Pcr ◽  
Virus Adsorption

scholarly journals Improved and Simplified Diagnosis of Covid-19 using TE Extraction from Dry Swabs

2020 ◽  
Author(s):  
Uday Kiran ◽  
C. G. Gokulan ◽  
Santosh Kumar Kuncha ◽  
Dhiviya Vedagiri ◽  
Karthik Bharadwaj Tallapaka ◽  
...  
Keyword(s):  
Mass Screening ◽  
Traditional Method ◽  
Reliable Method ◽  
Rna Isolation ◽  
New Method ◽  
Rt Pcr ◽  
Modified Method ◽  
Rigorous Testing ◽  
The Way

ABSTRACTRigorous testing is the way forward to fight the Covid-19 pandemic. Here we show that the currently used and most reliable RT-PCR based SARS-CoV-2 procedure can be further simplified to make it faster, safer and economical by bypassing the RNA isolation step. The modified method is not only fast and convenient but also at par with the traditional method in terms of accuracy, and therefore, can be used for mass screening. Our method takes about half the time and is cheaper by about 40% compared to current most widely used method. We also provide a variant of the new method that increases the efficiency of detection by about 20% compared to the currently used method. Taken together, we demonstrate a more effective and reliable method of SARS-CoV-2 detection.

scholarly journals Blockerette-Ligated Capture T7-Amplified RT-PCR, a New Method for Determining Flanking Sequences

2002 ◽  
Vol 6 (1) ◽  
pp. 113-118 ◽  
Author(s):  
Todd Lenvik ◽  
Troy C. Lund ◽  
Catherine M. Verfaillie
Keyword(s):  
New Method ◽  
Rt Pcr ◽  
Flanking Sequences

scholarly journals Simple Concentration Method Enables the Use of Gargle and Mouthwash Instead of Nasopharyngeal Swab Sampling for the Diagnosis of Covid19 by PCR

2021 ◽  
Author(s):  
Tanil Kocagoz ◽  
Ozge Can ◽  
Neval Yurttutan Uyar ◽  
Ece Aksoy ◽  
Tuba Polat ◽  
...  
Keyword(s):  
Sampling Procedure ◽  
New Method ◽  
Healthcare Personnel ◽  
Nasopharyngeal Swab ◽  
Test Results ◽  
Rt Pcr ◽  
Painful Procedure ◽  
Concentration Method ◽  
Before And After ◽  
Mouth Wash

Abstract Purpose Since its emergence in December 2019, SARS-CoV-2 is causing one of the most devastating pandemics in human history. Currently, the most important method for definitive diagnosis of COVID-19 is identification of SARS-CoV-2 RNA in nasopharyngeal swab samples by RT-PCR. Nasopharyngeal swab sampling is a discomforting procedure sometimes with adverse effects, which also poses a risk for infection for the personnel performing the sampling. We have developed a new method for concentrating biological samples, which enabled us to use gargle and mouth-wash samples to be used in RT-PCR, for the diagnosis of COVID-19, as an alternative to nasopharyngeal swab samples.Methods We have analyzed nasopharyngeal and gargle and mouthwash samples, before and after concentration, of 363 patients by RT-PCR for the presence of SARS-CoV-2.Results Among 114 patients in which SARS-CoV-2 was identified in at least one of their samples, the virus was identified in 76 (66.7%), 67 (58.8%) and 101 (88.6%) of nasopharyngeal swab, gargle and mouth-wash samples before and after concentration, respectively.Conclusion When concentrated by our new method, gargle and mouthwash samples can be used instead of nasopharyngeal samples in identification of SARS-CoV-2 by RT-PCR, with the same or better sensitivity. Eliminating the need for nasopharyngeal sampling, will save the patients from an invasive and painful procedure and will lower the risk of infection for the healthcare personnel taking the sample. This easy sampling procedure may decrease the workload of hospitals, shorten the turn-around time of obtaining test results and thus enable rapid isolation of infected patients.

Sign in / Sign up

Export Citation Format

Share Document