The suitability of using a modified real-time reverse transcriptase PCR method in the detection of Enterovirus in respiratory samples from Iraq

The Enterovirus is an important infectious agent that should focus on for better recognition and control. The current work was initiated to use a modified real-time reverse transcriptase PCR (RT-PCR) technique. For this aim, clinical specimens of300 human septum samples from patients with respiratory symptomswere obtained different regions in Al-Diwaniyah province from Iraq. The study was aimed at using VP1 gene based target with a specific primer set. The results of the RT-PCR method uncovered the presence of Enterovirus in 96(32%) samples. The method findings demonstrated successful rates with sensitivity (100%) and specificity (95%). The results provide evidence about the probability of the existence of the virus in those samples plus give critical importance about the use of this method in the identifying the target virus alone with high accuracy. Keyword: Enterovirus, RT-PCR, VP1 gene How to cite this article: Saihood AS, Saihood AS, Rayshan AR(2021): The suitability of using a modified real-time reverse transcriptase PCR method in the detection of Enterovirus in respiratory samples from Iraq, Ann Trop Med & Public Health; 24(S2): SP24237. DOI: http://doi.org/10.36295/ASRO.2021.24237 Introduction Enterovirus is one of the most common human pathogenic microorganisms, causing 10 to 15 million new infections in the United States per year. Therefore, both men have several EV diseases during their lives. There are 116 EV types in all four Enterovirus variants (A, B, C, D). Enteroviruses may induce either asymptomatic infections or acute diseases like diarrhea to encephalitis. Enterovirus disease, during early life years, offers lifetime immunity toward Saihood et al (2021): Use of reverse transcriptase PCR for detecting Enterovirus from respiratory samples Feb 2021 Vol. 24 Issue 2 Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2021.24237 post-homologous virus exposure and can also help to prohibit the occurrence of autoimmune diseases.Human Enterovirus was first identified from children with respiratory infections. The Fermon strain was referred to as the variant type obtained from samples of those children. However, till the beginning of the 2000s, there had not yet been a significant number of the virus occurrences. Since 2008 to 2012, nevertheless, the virus has been identified as an evolving microorganism sufficient to cause serious respiratory diseases in some of Asian, European, America countries such as the Philippines, Japan, and the Netherlands and the United States (1–4) . Initially categorized via serotyping, the genetic relatedness of entererovirus types which cause human infections is now categorized into 4 species (EV-A to EV-D). The group comprises three rhinovirus members (RV-A to RV-C, common cold triggers) and five animal affecting organisms. Most recently, Picornaviridae has produced a new family, the Parechoviruses, which included identical yet genetic variations, which were historically known as enteroviruses. Such viruses may trigger Enterovirus-like clinical diseases, although a large scale is not yet studied in conjunction with more chronic diseases (5–7) . The genomes of the Enterovirus are positive-sense RNA-single stranded with total length of 7.4 kilobases. It consist of a 5′′ (5′UTR) untranslated region, a single polyprotein coding area and 3′′ a small untranslated area with a 3′′ polyadenylated tail. The 5UTR is fairly well-maintained and is typically utilized for PCR research of medical sample identification. In the RNA replication regulation and polyprotein translation both 5′UTR and 3′UTR are engaged. This is then cleaved into eleven mature proteins, using proteases, (VP1toVP-4) which are the viral capsid, using an RNA polymerase manner throughout post-translation refining.The genes that encode functional proteins produce a high level of biodiversity and contribute to a wide range of different serotypes. The VP1 region's sequence demonstrates the most highly correlated methods of conventional serotyping, which are utilized most frequently in Enterovirus detection. Respiratory (upper respiratory Tract, URT) and Faecal-oral (gastrointestinal tract, GIT)routes play a major role in spreading enteroviruses. Incubation is usually only a few days, with replication happening primarily in the URT or in the GIT (via inhaling or exposure to oral or nasal mucosa) (8) . The Enterovirus is an important infectious agent that should focus on for better recognition and control. The current work was initiated to use a modified RT-PCR technique. Saihood et al (2021): Use of reverse transcriptase PCR for detecting Enterovirus from respiratory samples Feb 2021 Vol. 24 Issue 2 Annals of Tropical Medicine & Public Health http://doi.org/10.36295/ASRO.2021.24237 Materials and methods