Analytical Performance of COVID-19 Detection Methods (RT-PCR): Scientific and Societal Concerns

Abstract

Background. Health and social management of the SARS-CoV-2 epidemic, responsible for the COVID-19 disease, requires both screening tools and diagnostic procedures. Reliable screening tests aim at identifying (truely) infectious individuals that can spread the viral infection and therefore are essential for tracing and harnessing the epidemic diffusion. Instead, diagnostic tests should supplement clinical and radiological findings, thus helping in establishing the diagnosis. Several analytical assays, mostly using RT-PCR-based technologies, have become commercially available for healthcare workers and clinical laboratories. However, such tests showed some critical limitations, given that a relevant number of both false-positive and false-negative cases have been so far reported. Moreover, those analytical techniques demonstrated to be significantly influenced by pre-analytical biases, while the sensitivity showed a dramatic time dependency. Aim. Herein, we critically investigate limits and perspectives of currently available RT-PCR techniques, especially when referring to the required performances in providing reliable epidemiological and clinical information. Key Concepts. Current data cast doubt on the use of RT-PCR swabs as a screening procedure for tracing the evolution of the current SARS-COV-2 pandemic. Indeed, the huge number of both false-positive and false-negative results deprives the trustworthiness of decision making based on those data. Therefore, we should refine current available analytical tests to quickly identify individuals able to really transmit the virus, with the aim to control and prevent large outbreaks.

SARS-CoV-2: The Self-Nonself Issue and Diagnostic Tests

“…Clinically and of utmost importance in diagnostics, the viral versus human pentapeptide and oligonucleotide sharing shown in ►Table 1 and 2, respectively, could have a severe impact on the validity of the current polymerase chain reaction (PCR) tests for SARS-CoV-2 spike detection. In fact, claims have been reported about the rates of false negatives/positives in SARS-CoV-2 detection by means of serological and PCR tests, [33][34][35][36][37][38] in this way raising numerous concerns. As observed by Viswanathan et al, 39 healthy individuals may be falsely identified as positive, requiring confirmatory testing and potentially leading to the unnecessary isolation of these individuals.…”

Nucleotide Sequence Sharing between the Human Genome and Primers for Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Detection

“…However, especially in analyzing SARS-CoV-2 as a paradigmatic example, contrasting data have been reported on the analytical performance of SARS-CoV-2 detection methods and claims about the rates of false negatives and false positives have been published. 6 7 8 9 10 11…”

“…However, especially in analyzing SARS-CoV-2 as a paradigmatic example, contrasting data have been reported on the analytical performance of SARS-CoV-2 detection methods and claims about the rates of false negatives and false positives have been published. [6][7][8][9][10][11] On the basis of all these, this study focused on the possible genetic basis of potential false polymerase chain reaction (PCR) results by comparing the nucleotide sequence of proposed/used SARS-CoV-2 primers versus the human genome. The scientific rationale is that-given the high level of amino acid sequence sharing between SARS-CoV-2 proteins and the human proteome [12][13][14][15] -parallel sequence matching at the nucleotide level might exist between the SARS-CoV-2 primer sequences and the human genome, in this way possibly explaining the generation of false-positive SARS-CoV-2 detection results.…”

The use of salivary specimen for COVID-19 detection using RT-PCR assay: a systematic review

“…Active case finding through contact tracing and performing testing are mandatory to prevent mass transmission since many active cases are asymptomatic [14]- [16]. Several methods have been proposed to provide early detection of COVID-19, i.e., through detection of viral ribonukleat acid (RNA) [17], [18], viral antigen [19], [20], and serum antibody [21], [22]. Viral RNA detection using nucleic acid amplification test (NAAT) is known to have low limit of detection with excellence analytical specificity and among many different types of NAATs, reverse transcriptase Int J Public Health Sci ISSN: 2252-8806 …”