Diagnosis of SARS-CoV-2 during the Pandemic by Multiplex RT-rPCR hCoV Test: Future Perspectives

Abstract

Coronavirus disease 2019 (COVID-19), caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has rapidly become a significant threat to public health. However, among the Coronaviridae family members, there are other viruses that can also cause infections in humans. Among these, severe acute respiratory syndrome (SARS-CoV) and Middle East respiratory syndrome (MERS-CoV) have posed significant threats to human health in the past. Other human pathogenic coronaviruses have been identified, and they are known to cause respiratory diseases with manifestations ranging from mild to severe. In this study, we evaluated the performance of a multiplex RT-rPCR specific to seven human pathogenic coronaviruses in mainly detecting SARS-CoV-2 directly from nasopharyngeal swabs obtained from suspected COVID-19 infected patients, while simultaneously detecting different human pathogenic coronaviruses in case these were also present. We tested 1195 clinical samples suspected of COVID-19 infection. The assay identified that 69% of the samples tested positive for SARS-CoV-2 (1195), which was confirmed using another SARS-CoV-2 RT-PCR kit available in our laboratory. None of these clinical samples were positive for SARS-CoV, MERS-CoV or HCoV. This means that during the endemic phase of COVID-19, infection with other human pathogenic coronaviruses, even the common cold coronavirus (HCoV), is very uncommon. Our study also confirmed that the multiplex RT-rPCR is a sensitive assay for detecting SARS-CoV-2 regardless of differences among the variants. This multiplex RT-rPCR is also time- and cost-saving and very easy to apply in the diagnostic laboratory due to its simple procedure and its stability in storage after preparation. These features make the assay a valuable approach in screening procedures for the rapid detection of SARS-CoV-2 and other human pathogenic coronaviruses that could affect public health.

SARS-CoV-2-Specific T Cell Responses in Immunocompromised Individuals with Cancer, HIV or Solid Organ Transplants

“…The course of SARS-CoV-2 infection is variable and can be asymptomatic or may exhibit symptoms ranging from mild upper respiratory tract infection to multisystem organ failure and death [ 3 ]. A rapid diagnosis and differentiation from other common coronaviruses can be achieved with the application of tools such as multiplex PCR [ 4 , 5 , 6 ]. Additional parameters and techniques, including chest radiography-based evaluations of characteristic alterations in the lung, may further support the assessment of disease severity [ 3 , 7 , 8 ].…”

Multiplex RT Real-Time PCR Based on Target Failure to Detect and Identify Different Variants of SARS-CoV-2: A Feasible Method That Can Be Applied in Clinical Laboratories

“…The MPL RT-rPCR mixes of MPL1 and MPL2 were prepared by using one-step RT rPCR mix (AgPath-ID™ One-Step RT-PCR, Thermo, Waltham, MA, USA). Each mix contained 10 µL of Apath-ID RT-PCR buffer 2X, 0.8 µL of Apath-ID RT-PCR enzyme 25X, 10 pm of primers (2 pm for the RNAseP-specific primers), 5 pm of probes, 1 µL of enzyme stabilizer [ 5 , 6 ] (a substrance used to stabilize the enzyme in the prepared RT-rPCR mixes, which was supplied by Nam Khoa Co., Ltd., Ho Chi Minh City, Vietnam), and DNAse/RNAse-free DW at 15 µL. The MPL RT-rPCR mixes were pre-filled into low-profile, white PCR tubes suitable for the real-time PCR instrument used in this study (Biorad’s CFX-96) and kept at −20 °C or lower until used.…”

Damage from Carbonated Soft Drinks on Enamel: A Systematic Review

“…In addition, COVID-19, which saw a rise in unhealthy eating practices and a rise in the use of carbonated beverages, had a significant societal influence [ 13 , 14 ]. On December 12th, 2019, a new coronavirus (SARS-Cov2) emerged in China, sparking a pandemic of acute respiratory syndrome in humans, COVID-19 [ 15 , 16 , 17 ]. The COVID-19 pandemic represents a massive impact on human health, causing sudden lifestyle changes, through social distancing and isolation at home, with great social and economic consequences [ 18 , 19 , 20 ].…”

“…The consumption of soft drinks causes a reduction in the salivary pH [ 48 ]. A pH of 5.5 is considered the “critical pH” for enamel dissolution, while it is 6.8 for dentin [ 17 , 49 , 50 ]. Carbonated drinks have an extrinsic acidity and a pH of up to 2.5 because of the quantity of carbonic acid that is formed with the addition of CO2, which produces the fizz and other acids, such as citric acid, phosphoric acid, and tartaric acid [ 39 , 51 , 52 , 53 , 54 ].…”