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Quantification and Whole Genome Characterization of SARS-CoV-2 RNA in Wastewater and Air Samples
Published on: June 30, 2023
[SARS-CoV-2 and Microbiological Diagnostic Dynamics in COVID-19 Pandemic]
1Ege University Faculty of Medicine, Department of Medical Microbiology, Izmir, Turkey.
Insights
Understanding severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) dynamics is crucial for accurate Coronavirus Disease 2019 (COVID-19) diagnosis. Integrating RNA and antibody tests enhances diagnostic sensitivity, especially after the second week of illness.
Area of Science:
- Virology and Microbiology
- Infectious Diseases
- Immunology
Background:
- Coronavirus Disease 2019 (COVID-19), caused by SARS-CoV-2, presents high mortality and transmission rates.
- Understanding SARS-CoV-2's biological mechanisms, including host cell interaction via spike (S) protein and TMPRSS2, is vital for pandemic control.
- Accurate microbiological diagnosis is central to managing the COVID-19 pandemic and interpreting test results.
Purpose of the Study:
- To review current knowledge on SARS-CoV-2 and its microbiological diagnostic methods.
- To discuss the interpretation of diagnostic tests, including limitations and potential improvements.
- To highlight the role of integrated testing strategies for enhanced diagnostic accuracy.
Main Methods:
- Review of existing literature on SARS-CoV-2 virology and diagnostic techniques.
- Analysis of reverse-transcribed polymerase chain reaction (RT-PCR) for viral RNA detection.
- Evaluation of SARS-CoV-2 antibody tests (IgM, IgA, IgG) for immune response detection.
Main Results:
- RT-PCR detects viral RNA typically 2-3 days before symptom onset and in the first week; lower respiratory tract samples may be preferred later.
- Clinical sensitivity of SARS-CoV-2 RNA tests ranges from 55-75%, and RNA positivity does not confirm active infection.
- Antibody tests become detectable 5-14 days post-symptom onset, with potential for false positives in IgM and IgA tests due to low specificity.
Conclusions:
- Integrating SARS-CoV-2 RNA and antibody tests as co-tests significantly increases clinical sensitivity, particularly after the second week of illness.
- Negative RT-PCR results do not rule out SARS-CoV-2 infection.
- Further large-scale studies are needed to evaluate diagnostic performance metrics and seroconversion patterns across diverse populations.
Abstract:
We have been introduced to "Coronavirus Disease 2019 (COVID-19)" disease with high mortality and transmission rate caused by a novel human coronavirus, in December 2019 and the microbiological diagnosis of the infection has been in the center of the focus to control the pandemic. It is necessary to understand the dynamics of the virus which was classified among the severe acute respiratory syndrome (SARS) related coronaviruses and named as SARS coronavirus 2 (SARS-CoV-2), to manage testing in the right strategy and for interpretation of the results. However, much remains unclear about the virus and the immune response. SARS-CoV-2, which is an enveloped, RNA virus has been shown to attach to the host cell receptor angiotensin converting enzyme 2 with spike (S) protein and membrane fusion is provided by transmembrane protease serine 2 (TMPRSS2) of the host cell. The most commonly used and reliable test for diagnosis of COVID-19 is reverse-transcribed polymerase chain reaction (RT-PCR) performed by using nasopharyngeal swabs or other respiratory tract specimens. Viral RNA is usually detected two three days before the onset of symptoms and in the first week from upper respiratory tract samples. If possible, the lower respiratory tract specimens are preferable in the second week, especially if former PCR is negative and pneumonia has developed. The clinical sensitivity of SARS-CoV-2 RNA tests has been reported around 55-75%. Negative RT-PCR test result does not exclude COVID-19 or SARSCoV-2 infection. It should also be noted that viral RNA positivity is not an evidence of active or infectious virus. SARS-CoV-2 infection can be also detected indirectly by testing the host specific immune response to the virus. There is an increasing interest in the use of SARS-CoV-2 antibody tests both for the diagnosis and public health surveillance. However, the antibody tests should not be used as the sole test for diagnosis and case management. Antibody tests are valuable tools in seroepidemiological studies. Anti-SARS-CoV-2 IgM, IgA and IgG antibodies have been shown to be detectable as early as 5th-14th days after the onset of symptoms and most of them become positive on the 21st day. False positivity has been reported more frequently with IgM and IgA tests due to low specificity. It was shown that clinical sensitivity of the diagnostic approach increases when RNA and total antibody tests were integrated as co-tests, especially after the second week of the disease. Specificity, sensitivity, positive and negative predictive values are needed to be evaluated with large and standard studies targeting populations with different prevalences. It is also necessary to create evidence with larger seroconversion studies. In this review article, the information and data obtained until today about SARS-CoV-2 and its microbiological diagnosis have been discussed.

