We collected twenty-two (n = 22) nasopharyngeal samples (including COVID-19 = 8, Recovered = 7, and Healthy = 7) from Dhaka city of Bangladesh during May to July, 2020. The suspected patients were diagnosed positive for SARS-CoV-2 infections (COVID-19) through RT-qPCR. The confirmed patients were admitted into the dedicated COVID-19 isolation wards, and received medication. These patients were tested negative for COVID-19 after 17.5 (ranged from11 to 32) days of SARS-CoV-2 infection, and categorized as Recovered humans (S1 Table ). The nasopharyngeal swab samples from COVID-19 and Recovered subjects were collected on the test day (COVID-19 positive and COVID-19 negative confirmation day). The Healthy control subjects were randomly selected and these people did not show any signs and symptoms of respiratory illness. Nasopharyngeal swabs from these Healthy people were collected following the same protocol for COVID-19 and Recovered humans. The collected samples were placed in sample collection vial containing PBS (phosphate buffered saline). The RT-qPCR was performed for ORF1ab and N genes of SARS-CoV-2 using novel Coronavirus (2019-nCoV) Nucleic Acid Diagnostic Kit (PCR-Fluorescence Probing, Sansure Biotech Inc.) following the manufacturer’s instructions. The collected samples were immediately sent for RNA extraction, library preparation and sequencing [13 (link)].
2019 ncov nucleic acid diagnostic kit pcr fluorescence probing
Manufactured by Sansure Biotech
The (2019-nCoV) Nucleic Acid Diagnostic Kit (PCR-Fluorescence Probing) is a laboratory equipment product developed by Sansure Biotech. The kit is designed to detect the presence of the 2019 novel coronavirus (2019-nCoV) genetic material through real-time reverse transcription-polymerase chain reaction (RT-PCR) technology using fluorescence-labeled probes.
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Lab products found in correlation
2 protocols using 2019 ncov nucleic acid diagnostic kit pcr fluorescence probing
1
Nasopharyngeal Sampling for SARS-CoV-2 Detection
2
SARS-CoV-2 Detection via One-Step RT-PCR
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The collected air samples were transferred to a virology lab in cold chain and were kept at −70 °C. Total RNAs were extracted using an RNA Extraction Kit (Sinaclon Co, Tehran, IRAN). The purity of extracted RNAs was determined by Nanodrop (ThermoScientific, USA). One-step reverse transcriptase real-time PCR was performed for RNA-dependent RNA polymerase (RdRp) and N SARS-CoV-2 genes with specific primers and probes (FAM and Texas red) with the use of a One Step Novel Coronavirus (2019-NCOV) Nucleic Acid Diagnostic Kit (PCR-Fluorescence Probing, SANSURE Biotech, china). The internal control primer and probe (CY5) were evaluated. Thermal cycling conditions were as follows: 50 °C for 30 min, 95 °C for 1 min, followed by 45 cycles of 95 °C for 15 s, 60 °C for 30 s. The RT-PCR cycle threshold (CT) values ≤ 40 and a sigmoidal curve were considered as positive results for both genes. The sampling and extraction were repeated for samples in which the Ct of the Internal Control gene was not determined.
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