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Sensitivity of RT-PCR tests for SARS-CoV-2 through time.

ABSTRACT: Reverse transcriptase polymerase chain reaction (RT-PCR) tests are the gold standard for detecting recent infection with SARS-CoV-2. RT-PCR sensitivity varies over the course of an individual's infection, related to changes in viral load. Differences in testing methods, and individual-level variables such as age, may also affect sensitivity. Using data from New Zealand, we estimate the time-varying sensitivity of SARS-CoV-2 RT-PCR under varying temporal, biological and demographic factors. Sensitivity peaks 4-5 days post-infection at 92.7% [91.4%, 94.0%] and remains over 88% between 5 and 14 days post-infection. After the peak, sensitivity declined more rapidly in vaccinated cases compared to unvaccinated, females compared to males, those aged under 40 compared to over 40 s, and Pacific peoples compared to other ethnicities. RT-PCR remains a sensitive technique and has been an effective tool in New Zealand's border and post-border measures to control COVID-19. Our results inform model parameters and decisions concerning routine testing frequency.

SUBMITTER: Binny RN

PROVIDER: S-EPMC9384503 | biostudies-literature | 2022 Jul

REPOSITORIES: biostudies-literature

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Sensitivity of Reverse Transcription Polymerase Chain Reaction Tests for Severe Acute Respiratory Syndrome Coronavirus 2 Through Time.

Binny Rachelle N RN Priest Patricia P French Nigel P NP Parry Matthew M Lustig Audrey A Hendy Shaun C SC Maclaren Oliver J OJ Ridings Kannan M KM Steyn Nicholas N Vattiato Giorgia G Plank Michael J MJ

The Journal of infectious diseases 20221201 1

<h4>Background</h4>Reverse transcription polymerase chain reaction (RT-PCR) tests are the gold standard for detecting recent infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Reverse transcription PCR sensitivity varies over the course of an individual's infection, related to changes in viral load. Differences in testing methods, and individual-level variables such as age, may also affect sensitivity.<h4>Methods</h4>Using data from New Zealand, we estimate the time-var ...[more]

PMID: 35876500

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Project description:BackgroundSevere acute respiratory syndrome coronavirus 2 (SARS-CoV-2) which emerged in the city of Wuhan, Hubei Province, China, has spread worldwide and is threatening human life. The detection of SARS-CoV-2 is critical for preventing new outbreaks, curbing disease spread, and managing patients. Currently, a reverse-transcription polymerase chain reaction (RT-PCR) assay is used to detect the virus in clinical laboratories. However, although this assay is considered to have high specificity, its sensitivity is reportedly as low as 60-70 %. Improved sensitivity is, therefore, urgently required.MethodsWe used the primers and single-quencher probes recommended by the CDC (N1, N2 and N3) in the USA and the NIID (N1 and N2) in Japan. In addition, we designed double-quencher probes according to the virus sequence provided by the NIID to develop a further assay (termed the YCH assay [N1 and N2]). Using these assays, we conducted RT-PCR with serially diluted DNA positive controls to assess and compare the detection sensitivity of the three assays. Furthermore, 66 nasopharyngeal swabs were tested to determine the diagnostic performances.ResultsThe threshold cycle (Ct) value of the RT-PCR was relatively low for the CDC and YCH assays compared with the NIID assay. Serial dilution assays showed that both the CDC and YCH assays could detect low copy numbers of the DNA positive control. The background fluorescence signal at the baseline was lower for the YCH assay compared with the NIID assay. We assessed the diagnostic performance between single- (NIID) and double-quencher (YCH) probes using 66 nasopharyngeal swabs. When the results of YCH-N2 assay were used as a reference, each assay detected SARS-CoV-2 with positive percent agreements of 56 % for NIID-N1, 61 % for YCH-N1, and 94 % for NIID-N2, and 100 % negative percent agreements for NIID-N1, YCH-N1 and NIID-N2.ConclusionDouble-quencher probes decreased the background fluorescence and improved the detection sensitivity of RT-PCR for SARS-CoV-2.

Dataset Information

Sensitivity of RT-PCR tests for SARS-CoV-2 through time.

Publications

Sensitivity of Reverse Transcription Polymerase Chain Reaction Tests for Severe Acute Respiratory Syndrome Coronavirus 2 Through Time.

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