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1.
J AOAC Int ; 104(4): 872-888, 2021 Aug 20.
Article in English | MEDLINE | ID: covidwho-1387921

ABSTRACT

BACKGROUND: The Eurofins GeneScan Technologies' VIRSeek SARS-CoV-2 Mplex kit is a RT (reverse transcription) real-time polymerase chain reaction (RT-qPCR) assay for the detection of two targets on the N-gene (nucleocapsid) of SARS-CoV-2. An extraction control, that allows monitoring of the extraction procedure and PCR inhibition, is included. OBJECTIVE: In silico analysis and wet testing showed inclusivity and exclusivity of the assay. The complete workflow starting from surface swabbing (VIRSeek PATHOSwab kit), RNA extraction (VIRSeek RNAExtractor), RT-PCR (VIRSeek SARS-CoV-2 Mplex), and evaluation with FastFinder was validated in comparison to the CDC method for detection of SARS-CoV-2 on stainless steel. METHOD: In silico analysis was performed by using the MFOLD online program. The matrix study was performed for stainless steel inoculated with SARS-CoV-2 isolated from the first documented US case of a traveler from Wuhan, China. RESULTS: For inclusivity, 15 764 sequences were analyzed and all mismatches (0.37% of the sequences had single mismatches) were considered non-critical. Cross reactivity for closely related viruses and background organisms was performed, resulting in correct exclusion of all. No significant differences were observed for the probability of detection (POD) study when comparing to the CDC method. CONCLUSIONS: Results of the inclusivity and exclusivity study show that the assay is specific for detection of SARS-CoV-2. The POD study showed no statistically significant difference compared to the CDC reference method, results were identical for the uninoculated and the high level. For the fractional recovery level, the candidate method detected 9/17 samples leading to a POD of 0.47, the reference method detected 11/20 samples leading to a POD of 0.55. HIGHLIGHT: The complete workflow starting from swabbing of the surface (VIRSeek PATHOSwab kit), RNA extraction (VIRSeek RNAExtractor), RT-PCR (VIRSeek SARS CoV-2 Mplex) and evaluation with FastFinder was validated in comparison to the US Centers for Disease Control and Prevention method for detection of SARS-CoV-2 on Stainless Steel.


Subject(s)
COVID-19 , SARS-CoV-2 , Humans , RNA, Viral/genetics , Real-Time Polymerase Chain Reaction , Sensitivity and Specificity , Stainless Steel
2.
Transfus Apher Sci ; 59(6): 102922, 2020 Dec.
Article in English | MEDLINE | ID: covidwho-728868

ABSTRACT

SARS-CoV-2 has infected millions worldwide. The virus is novel, and currently there is no approved treatment. Convalescent plasma may offer a treatment option. We evaluated trends of IgM/IgG antibodies/plasma viral load in donors and recipients of convalescent plasma. 114/139 (82 %) donors had positive IgG antibodies. 46/114 donors tested positive a second time by NP swab. Among those retested, the median IgG declined (p < 0.01) between tests. 25/139 donors with confirmed SARS-CoV-2 were negative for IgG antibodies. This suggests that having had the infection does not necessarily convey immunity, or there is a short duration of immunity associated with a decline in antibodies. Plasma viral load obtained on 35/39 plasma recipients showed 22 (62.9 %) had non-detectable levels on average 14.5 days from positive test versus 6.2 days in those with detectable levels (p < 0.01). There was a relationship between IgG and viral load. IgG was higher in those with non-detectable viral loads. There was no relationship between viral load and blood type (p = 0.87) or death (0.80). Recipients with detectable viral load had lower IgG levels; there was no relationship between viral load, blood type or death.


Subject(s)
Antibodies, Viral/administration & dosage , COVID-19/blood , COVID-19/therapy , SARS-CoV-2 , Adult , Aged , Female , Humans , Immunization, Passive , Immunoglobulin G/administration & dosage , Immunoglobulin M/administration & dosage , Male , Middle Aged
3.
J Clin Virol ; 129: 104439, 2020 08.
Article in English | MEDLINE | ID: covidwho-306092

ABSTRACT

Real-time reverse transcriptase polymerase chain reaction (rRTPCR) has been the main method for diagnosis of SARS-CoV-2 infection in the early stages of the COVID-19 pandemic. De-identified results from upper and lower respiratory samples submitted to a reference laboratory demonstrated a positivity rate of 14.9 % (4428 of 29,713 samples tested). Distribution of results by birth year cohort and specimen type suggested general consistency in mean, median and peak values but higher positivity rates in individuals born from 1964 to 1974. Female patients had a significantly lower positivity rate (P < 0.0001), although similar load mean and median values, compared to males. Overall, 15.3 % (676 of 4428 positive results) of positive results had viral loads greater than 8 log10 copies/mL, with occasional samples exceeding 10 log10 copies/mL. These results support quantitative assessment of SARS-CoV-2 viral load in patient testing and efforts to control viral transmission.


Subject(s)
Betacoronavirus/isolation & purification , Clinical Laboratory Techniques , Coronavirus Infections/virology , Pneumonia, Viral/virology , Respiratory System/virology , Viral Load , Adult , Age Factors , Aged , Aged, 80 and over , COVID-19 , COVID-19 Testing , COVID-19 Vaccines , Child , Child, Preschool , Coronavirus Infections/diagnosis , Female , Humans , Infant , Infant, Newborn , Male , Middle Aged , Pandemics , SARS-CoV-2 , Sex Factors , Young Adult
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