Estimation of primer efficiency in multiplex PCR for detecting SARS-Cov-2 variants
Lubna Muhi Rasoul, Mohanad M. Nsaif, Mohammed Hamid Habeeb AL-Tameemi, Bahaa Abdullah Laftaah Al-Rubaii
Abstract
In periodontology, polymerase chain reaction (PCR) has emerged as a standard diagnostic and research instrument. According to the findings in research, its sensitivity and specificity make it an efficient and speedy method for detecting, identifying, and quantifying organisms. The spike (S) gene, the envelope (E) gene, the membrane (M) protein, the nucleocapsids (N) gene, and the open reading frame (ORF) were used for the molecular identification of various variations of SARS-Cov-2 infection since they are critical components in the viral genome. Using kits approved by the Iraqi Ministry of Health following the spread of the covid-19 pandemic, this study sought to shed light on the significance of primers in the identification of various SARS-COV-2 variations involving many genes such as spike (S), envelope (E), membrane (M) protein, nucleocapsid (N), and ORFab1. A total of (150) clinically diagnosed patients for COVID-19 were screened for this study to estimate the accuracy and efficacy of primers used in detecting these genes are precise in diagnosing the infection in correspondence with clinical findings. The findings revealed that 18 (22.666%) of the cases had severe symptoms such as fever, unproductive hacking, breathlessness, muscle aches, and weariness, Whereas 42 (28%) of patients had lesser symptoms, and 74 (49.333%) of cases had minor signs on their own.. Results showed that all genes were accurate and efficient in the molecular diagnosis of the virus during the period from the beginning of March 2021 up to the writing of this manuscript in December 2021; the conclusion indicates that primers used in detecting these genes are efficient and accurate in the diagnosis of different variations of SARS-CAV-2 and there is no alteration in the viral genome in those genes that can disturb the result. Keywords: SARS-Cov-2 variations, spike (S) gene, envelope (E) gene, membrane (M) protein, Nucleocapsids (N), multiplex PCR