SARS-CoV-2 Detection using Real Time PCR by a Commercial Diagnostic Kit

Area of Science:

• * Medical Microbiology
• * Virology
• * Public Health

Background:

• * The emergence of SARS-CoV-2 caused a global public health crisis, leading to COVID-19.
• * Limited virus isolates presented challenges for public laboratories in accurate SARS-CoV-2 detection.
• * Early outbreak assessments indicated a wider spread than initially perceived.

Purpose of the Study:

• * To evaluate the diagnostic performance of a specific real-time PCR kit for SARS-CoV-2 detection.
• * To assess the kit's effectiveness in a clinical laboratory setting in Bangladesh.

Main Methods:

• * A Novel Corona virus (2019-nCoV) Nucleic Acid Diagnostic Kit (PCR-Fluorescence Probing), a real-time reverse transcription polymerase chain reaction (rRT-PCR) test, was utilized.
• * A total of 230 samples were analyzed from the microbiology department of Mymensingh Medical College.
• * Detection of ORF 1ab, N gene, and IC gene was performed using specific fluorescent dyes (FAM, ROX, CY5).

Main Results:

• * Out of 230 samples, 20 (8.69%) tested positive for SARS-CoV-2.
• * Among positive samples, 55% amplified both ORF 1ab and N genes; 15% and 30% amplified only ORF 1ab or N gene, respectively.
• * High amplification rates were observed for ORF 1ab (92.8%) and N gene (100%) with Ct values <38. Internal Control (IC) gene showed 70% positivity in positives and 100% in negatives.

Conclusions:

• * The study kit demonstrated significant capacity for detecting SARS-CoV-2 in laboratory settings.
• * The rRT-PCR diagnostic kit is a reliable tool for identifying SARS-CoV-2 in Bangladesh.
• * The findings support the use of this kit for routine diagnostics during the COVID-19 pandemic.