scholarly journals Adjusting the Cut-Off and Maximum Pool Size in RT-qPCR Pool Testing for SARS-CoV-2

Viruses ◽  
2021 ◽  
Vol 13 (4) ◽  
pp. 557
Author(s):  
Murilo S. Costa ◽  
Hugo I. Sato ◽  
Raissa P. Rocha ◽  
Alex F. Carvalho ◽  
Nathalia S. Guimarães ◽  
...  
Keyword(s):  
Large Scale ◽  
Pool Size ◽  
Individual Sample ◽  
Chain Reaction ◽  
Test Sensitivity ◽  
Large Scale Testing ◽  
Public Emergency ◽  
Polymerase Chain ◽  
Belo Horizonte

Reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) to detect SARS-CoV-2 RNA is an essential test to monitor the occurrence of COVID-19. A methodology is proposed for the determination of maximum pool size and adjustments of cut-off values of cycle threshold (Ct in RT-qPCR pool testing, to compensate for the dilution caused by pooling. The trade-off between pool size and test sensitivity is stated explicitly. The procedure was designed to ensure that samples that would be detectable in individual testing remain detectable in pool testing. The proposed relaxation in cut-off is dependent on the pool size, allowing a relatively tight correction to avoid loss of detection of positive samples. The methodology was evaluated in a study of pool testing of adults attending a public emergency care unit, reference for COVID-19 in Belo Horizonte, Brazil, and presenting flu-like symptoms. Even samples on the edge of detectability in individual testing were detected correctly. The proposed procedure enhances the consistency of RT-qPCR pool testing by enforcing that the scales of detectability in pool processing and in individual sample processing are compatible. This may enhance the contribution of pool testing to large-scale testing for COVID-19.

Determination of N-myc gene amplification in neuroblastoma by differential polymerase chain reaction

1993 ◽  
Vol 7 (3) ◽  
pp. 227-234 ◽  
Author(s):  
J. Gilbert ◽  
M.D. Norris ◽  
M. Haber ◽  
M. Kavallaris ◽  
G.M. Marshall ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Gene Amplification ◽  
Chain Reaction ◽  
Myc Gene ◽  
Polymerase Chain

Determination of a T/G polymorphism at nucleotide 3206 of the apolipoprotein C III gene by amplification refractory mutation system

1994 ◽  
Vol 40 (12) ◽  
pp. 2235-2239 ◽  
Author(s):  
M Y Tsai ◽  
N Q Hanson ◽  
K R Copeland ◽  
I Beheshti ◽  
U Garg
Keyword(s):  
Epidemiological Studies ◽  
Amplification Refractory Mutation System ◽  
Chain Reaction ◽  
Apolipoprotein C ◽  
Significant Difference ◽  
Mutation System ◽  
Polymerase Chain ◽  
Exon 4 ◽  
And Control

Abstract We used the amplification refractory mutation system (ARMS)--a polymerase-chain-reaction-based method--to determine the 3206 T-to-G polymorphism on exon 4 of the apolipoprotein (apo) C III gene. Apo C III is an inhibitor of the enzyme lipoprotein lipase (EC 3.1.1.34). Previous studies have demonstrated that a polymorphism at nucleotide 3175 on exon 4 of this gene is associated with hypertriglyceridemia. We studied 45 hypertriglyceridemic and 46 age-matched controls for the 3206 T-to-G polymorphism. The results showed a significant difference in the distribution of the genotypes with respect to this allele between the hypertriglyceridemic and control individuals. We also determined the presence of the SacI site at nucleotide 3175 in these same individuals and found no significant difference in SacI genotypes between the two groups. This study reaffirms the usefulness of ARMS as a simple, reliable method for detecting mutations and polymorphisms in clinical and epidemiological studies.

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