Molecular diagnostic technologies for COVID-19: Limitations and challenges

Afzal, Adeel

doi:10.1016/j.jare.2020.08.002

Cited by 290 publications

(280 citation statements)

References 82 publications

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“…However, the analytical specificity of the LDT is reported as 100% [ 19 ] and it demonstrated a high negative percent agreement with the CDC N1/N2 assay. Most other SARS-CoV-2 rtRT-PCR assays have shown high clinical specificities, making this a reasonable assumption [ 26 , 27 ].…”

Section: Discussionmentioning

confidence: 99%

False negative rate of COVID-19 PCR testing: a discordant testing analysis

Kanji

Zelyas

MacDonald

et al. 2021

Virol J

181

151

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Background COVID-19 is diagnosed via detection of SARS-CoV-2 RNA using real time reverse-transcriptase polymerase chain reaction (rtRT-PCR). Performance of many SARS-CoV-2 rtRT-PCR assays is not entirely known due to the lack of a gold standard. We sought to evaluate the false negative rate (FNR) and sensitivity of our laboratory-developed SARS-CoV-2 rtRT-PCR targeting the envelope (E) and RNA-dependent RNA-polymerase (RdRp) genes. Methods SARS-CoV-2 rtRT-PCR results at the Public Health Laboratory (Alberta, Canada) from January 21 to April 18, 2020 were reviewed to identify patients with an initial negative rtRT-PCR followed by a positive result on repeat testing within 14 days (defined as discordant results). Negative samples from these discordant specimens were re-tested using three alternate rtRT-PCR assays (targeting the E gene and N1/N2 regions of the nucleocapsid genes) to assess for false negative (FN) results. Results During the time period specified, 95,919 patients (100,001 samples) were tested for SARS-CoV-2. Of these, 49 patients were found to have discordant results including 49 positive and 52 negative swabs. Repeat testing of 52 negative swabs found five FNs (from five separate patients). Assuming 100% specificity of the diagnostic assay, the FNR and sensitivity in this group of patients with discordant testing was 9.3% (95% CI 1.5–17.0%) and 90.7% (95% CI 82.6–98.9%) respectively. Conclusions Studies to understand the FNR of routinely used assays are important to confirm adequate clinical performance. In this study, most FN results were due to low amounts of SARS-CoV-2 virus concentrations in patients with multiple specimens collected during different stages of infection. Post-test clinical evaluation of each patient is advised to ensure that rtRT-PCR results are not the only factor in excluding COVID-19.

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Section: Discussionmentioning

confidence: 99%

False negative rate of COVID-19 PCR testing: a discordant testing analysis

Kanji

Zelyas

MacDonald

et al. 2021

Virol J

181

151

View full text Add to dashboard Cite

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“…RT-PCR is a sensitive and rapid detection tool in molecular diagnostics. It can detect and amplify even a few copies of specific genomic sequence in a variety of samples, but it depends upon certain aspects to deliver reliable results like proper collection, transport, storage, and processing of samples (Afzal, 2020). It has been used for detection of diverse viruses like Adenovirus, Rotavirus, Astroviruses and many enteric viruses isolated from fecal samples (Kowada et al, 2018).…”

Section: Rt-pcr As a Frontline Diagnostic Methods For Covid-19 Diagnosismentioning

confidence: 99%

“…They developed an assay kit, which was commercialized as TaqPath™ COVID-19 combo kit. This strategy lowered the risk of contracting infection during sample collection by the health practitioners (Afzal, 2020). Sample collection for protein-based diagnosis like IgG/IgM and LFA, requires patients' blood samples.…”

Section: Sample Collection and Biosafety Measuresmentioning

confidence: 99%

An Update on Molecular Diagnostics for COVID-19

Islam

Iqbal

2020

Front. Cell. Infect. Microbiol.

101

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“…However, the search for class-switched isotypes, in this case IgG, might help to decrease the risk of errors. In addition, a target antigen is also essential when the virus being tested is capable of mutating because the same viral antigens will be present in its structure (93).…”

Section: Enzyme Linked Immunosorbent Assay (Elisa)mentioning

confidence: 99%

Trends in MERS-CoV, SARS-CoV, and SARS-CoV-2 (COVID-19) Diagnosis Strategies: A Patent Review

Nascimento

Santos

Oliveira

et al. 2020

Front. Public Health

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The emergence of a new coronavirus (SARS-CoV-2) outbreak represents a challenge for the diagnostic laboratories responsible for developing test kits to identify those infected with SARS-CoV-2. Methods with rapid and accurate detection are essential to control the sources of infection, to prevent the spread of the disease and to assist decision-making by public health managers. Currently, there is a wide variety of tests available with different detection methodologies, levels of specificity and sensitivity, detection time, and with an extensive range of prices. This review therefore aimed to conduct a patent search in relation to tests for the detection of SARS-CoV, MERS-CoV, and SARS-CoV-2. The greatest number of patents identified in the search were registered between 2003 and 2011, being mainly deposited by China, the Republic of Korea, and the United States. Most of the patents used the existing RT-PCR, ELISA, and isothermal amplification methods to develop simple, sensitive, precise, easy to use, low-cost tests that reduced false-negative or false-positive results. The findings of this patent search show that an increasing number of materials and diagnostic tests for the coronavirus are being produced to identify infected individuals and combat the growth of the current pandemic; however, there is still a question in relation to the reliability of the results of these tests.

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Molecular diagnostic technologies for COVID-19: Limitations and challenges

Abstract: Graphical abstract

Cited by 290 publications

References 82 publications

False negative rate of COVID-19 PCR testing: a discordant testing analysis

False negative rate of COVID-19 PCR testing: a discordant testing analysis

An Update on Molecular Diagnostics for COVID-19

Trends in MERS-CoV, SARS-CoV, and SARS-CoV-2 (COVID-19) Diagnosis Strategies: A Patent Review

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