Shallal et al (2020): Prevalence of COVID-19 in Raparin July 2020 Vol. 23 Issue 11

Prevalence of SARS-CoV-2 (COVID-19) in Raparin district Ahmed Farhan Shallal*1, Omed Hassan Qadir2, Ramiar Kamal Kheder1,Jangy Esmail Abdulla3,Jamal K. Shakor4, Rawand Abdulrahman Isa2

1Medical Laboratory Science department, College of Science, University of Raparin, , KGR, 2Ministry of health, Regional Government of Iraq, General Ranya Hospital 3College of Nursing, University of Raparin, Ranya, KGR, Iraq 4Department of Nursing, Sulaimani Polytechnic University, Iraq

*Corresponding Author: [email protected] (Shallal)

ABSTRACT Background: The COVID- 19 disease linked to the severe acute respiratory syndrome coronavirus virus 2 (SARS- CoV-2), it is a pandemic virus which has high fatality rate. Social gathering is being proposed as a risk factor for the disease. The aim of this study was to find out the level of Coronavirus (COVID-19) infection in Raparin district/Sulaimania governorate/ KGR, Iraq.

Materialsand Methods: The Five hundred and two nasopharyngeal swab samples were collected from different screening programsduring the second and third month of the year 2020. The samples randomly were collected from people who are in high-risk placesfor spreading COVID-19 such as petrol pump stations, markets, and pharmacy stores. Nuclear material was extracted using the PowerPrepTMViral DNA/RNA Extraction kit(E0007). Real time PCR was used using PowerChekTM2019-nCoV Real-time PCR kit.

Results: The current studydemonstrated that the mean age of the screened population was 34.45±12.55,the majority of whom were male (81.7). The screened population was mostlyfrom (68. 3%),while other samples were tested from the people who came back from other countries(20.2%). Overall surveyed people were asymptomatic and not presented with other morbidities. The prevalence of COVID-19 among the surveyed population was 4/1000 population.

Conclusions:Through the results of the current study, our conclusion wasthat quarantine and social separation, in addition to the curfew, are the main factors in preventing and spreading infection with Coronavirus.

Keywords: COVID-19, Quarantine,Viral Infection, Real-time RT-PCR

How to cite this article: Shallal AF, et al (2020):Prevalence of SARS-CoV-2 (COVID-19) in Raparin district, Ann Trop Med & Public Health; 23(S11): SP23117. DOI: http://doi.org/10.36295/ASRO.2020.231107

INTRODUCTION The emerging of coronavirus has become a global issue in the world. Companies and researches at universities are trying to develop various types of a vaccine against the coronavirus; some of them have been tested on human as a trial [1,2]. Coronaviruses are a group of viruses belonging to the Coronaviridae family in the Nidovirales order [3]. Coronaviruspossesses crown-like spikes on the outer surface of the virus; therefore, it was named as a coronavirus. Coronaviruses are, small in size (65–125 nm in diameter), enveloped, non-segmented, positive‐sense single‐stranded RNA virus genomes in the size ranging from 26 to 32 kilobases in length, the largest named viral RNA genome. Coronavirus can be divided into different subgroups including family alpha (α), beta (β), gamma (γ) and delta (δ) coronavirus [4-6].

The severe acute respiratory syndrome coronavirus (SARS-CoV), H5N1 influenza A, H1N1 2009 and Middle East respiratory syndrome coronavirus (MERS-CoV) are developed acute lung injury and acute respiratory distress

Annals of Tropical Medicine & Public http://doi.org/10.36295/ASRO.2020.231107

Shallal et al (2020): Prevalence of COVID-19 in Raparin July 2020 Vol. 23 Issue 11 syndrome which causes pulmonary failure and fatality in severe cases. It was known that these viruses infect only animals until the world suffered a severe acute respiratory syndrome (SARS) outbreak as a result of SARS-CoV, 2002 in Guangdong, China [7,8]. After 10 years, another pathogenic coronavirus, called Middle East respiratory syndrome coronavirus (MERS-CoV) resulted in an endemic in Middle Eastern countries. At the end of 2019, at Wuhan city, there was an outbreak of deadly coronavirus, the name of the coronavirus calledSARS-CoV-2 virus, which causes coronavirus disease-2019 (COVID-19), can cause lung complications such as pneumonia, and in most severe cases lead to other organ failures such as kidney and death [8-10]. This virus was known to be a member of the β group of coronaviruses. Chinese researchers named the virus as Wuhan coronavirus or 2019 novel coronavirus (2019-nCov). SARS-CoV-2 virus, COVID-19 was named by the International Committee on Taxonomy of Viruses [11-13]. The novel coronavirus (2019) infected 3,778,012individuals with a mortality rate of 3.4 %, across 212 countries, untilthe date of this writing.

There was evidence that bats are the major emerging sources of SARS-CoV because Chinese horseshoe bats include serologic evidence for prior infection with a related CoV and they possess sequences of SARS-related CoVs[14- 16].The proper evidence that SARS-CoV originated in bats was that they had the same receptor as the human virus, angiotensin-converting enzyme 2 (ACE2)[17].Although the basic outlines of disease transmission have not been ended by COVID-19,because it is a respiratory disease virus, it transmits mainly by direct contact of infected individuals with healthy people, itdevelops due to close contact with an infected individual, exposed to respiratory droplets, aerosols, coughing and sneezing (18, 19). These aerosols can enter the lungs of the human body through inhalation of the nose or mouth. There are some ways to protect people from spreading coronavirus infection, the main method is controlled through the use of quarantining [18,19].This research was performed on people in Raparindistrict in Kurdistan region of Iraq, the aim was to investigate the prevalence of corona virus in Raparindistrict/ Rania- Sulaimnia- Kurdistan region-Iraq.

MATERIALS AND METHODS Subjects Five hundred and two nasopharyngeal swab samples were collected from people at General Ranya Hospital in Sulymanya governorate (Molecular virology Lab)to detect SARS-CoV-2 by real-time reverse transcriptase (RT)- PCR. RNA was extracted from clinical samples with a PowerPrepTM Viral DNA/RNA Extraction kit (Kogenebiotech, Korea) following the manufacturer’s instructions. All samples were prepared under a biosafety cabinet according to laboratory biosafety guidelines. The viruses were diluted with viral transfer medium (VMT).The real-time PCR was perfumed using PowerChekTM2019-nCoV Real-time PCR kit.

Health survey

The work was conducted over various screening programs during the period February 2020 March 2020. The samples were collected from people who had a history of traveling outside of Kurdistan region. In addition, the cases were collected from peoples who areat high-risk for spreading COVID-19such as petrol pump stations, markets, and pharmacy stores. The result had only one positive forCOVID-19 out of 502 samples,the positive case was a worker in a petrol pump station who had history contact with a traveler from Iran country.

Real-time RT-PCR PowerPrepTMViral DNA/RNA Extraction kit(E0007) was used for purification of viral RNA from swab samples. The Kit includes all reagents necessary for the purification of viral nucleic acid from collected samples (Table1).

Table (1) shows the kit components for viral RNA extraction Components Volume Quantity Storage Lysis Buffer 30 ml 1 bottle Binding Buffer 40ml 1 bottle Wash Buffer A 30ml 1 bottle Room temperature Wash Buffer B 30ml 1 bottle Elution Buffer 15ml 1 bottle Spin Columns - 50 columns Micro-centrifuge tube - 50 tubes

Annals of Tropical Medicine & Public http://doi.org/10.36295/ASRO.2020.231107

Shallal et al (2020): Prevalence of COVID-19 in Raparin July 2020 Vol. 23 Issue 11

PowerChekTM 2019-nCoV Real-time PCR kit provides the fast and accurate testing solution for Wuhan coronavirus, specifically targeting the E gene for beta coronavirus and RdRp gene for 2019-n Cov in nasopharyngeal swab samples. This assay was based on the WHO and KCDC reference method (Table2).

Table (2) shows the kit components for detection Beta- coronavirus and 2019-nCoV

Component parts Cap Volume RT-PCR RP 1.100 µl Primer/ Probe Mix1 (E gene) P1 200 µl Primer/ Probe Mix2 (RdRp gene) P2 200 µl Control 1 (E gene) C1 50 µl Control 2 (RdRp gene) C2 50 µl

PCR Mixture The total reaction volume was 20 µl as shown in thetable (3).The reverse transcription process was performed at 50°C for 30 minutes, and then followed by inactivation of the reverse transcriptase enzyme at 95°C for 10 minutes. PCR amplification process was performed with 40 cycles at 95°C for 15 seconds and 60°C for 1 minute using an R6900T[20, 21].

Components Volume (µl) RT-PCR premix 11 Each primer/ probe Mix 4 Template RNA X(control 5) Total 20

Statistical analysis Statistical Package for Social Science (SPSS) V20 was used to analyze a descriptive analysis such as frequency and percentage. RESULTS

The characteristic of the quarantine population and COVID-19 results are given in (Table 4). Table (4) Characteristic of quarantine population and COVID-19 results Variables Frequency Percentage Gender Female 87 17.3 Male 410 81.7 Total 502 100.0 Mean of age 34.45±12.55 Place where lived

Annals of Tropical Medicine & Public http://doi.org/10.36295/ASRO.2020.231107

Shallal et al (2020): Prevalence of COVID-19 in Raparin July 2020 Vol. 23 Issue 11

European countries 102 20.2 US and Canada 6 1.1 Iran 50 9.9 Azerbaijan 1 0.01 Kurdistan/ Raparin Region 343 68.3 Cough Yes 0 0 No 502 100 Dyspnea Yes 0 0 No 502 100 Fever Yes 1 0.001 No 501 99.99 Cold Yes 0 0 No 502 100 Medical History Yes 1 0.001 No 502 99.99 COVID-19 Results Yes 2 0.004 No 500 99.8

DISCUSSION COVID-19 pandemic is one of the deadliest and most severe diseases of the twenty-first century, due to the health catastrophe it has caused around the world. Strict infection control, patient isolation and quarantineare influential in limiting the infecting and spreading[22]. Five hundred and two nasopharyngeal swab samples were collected from susceptible people in the Raparin region in the Sulaimania governorate. The screening was performed for people who came back from European countries (20.2%) and those who had contact with them (68. 3%), and the overall people were asymptomatic. The mean age was 34.45±12.55, majority of them were male 81.7%. The prevalence of COVID-19 among the surveyed population was 4/1000 population. One of the cases was detected from the traveler screening group and the other was from high-risk place screening group. The screening programs have a distinct detection rate and efficacy. A study has proved that a traveler screening program could effectively decrease the spread of COVID-19 but could not limit [23]. The positive rate or prevalence rate of the COVID-19 case among the screened population is mostly related to people who are targeted to screening. In chines study, among those who had contact with COVID-19 cases or had respiratory tract infection’s symptoms, the positive rate was 38% [24].Vaccination with BCG less infected with coronavirus, because this vaccine trained immune system to recognize Annals of Tropical Medicine & Public http://doi.org/10.36295/ASRO.2020.231107

Shallal et al (2020): Prevalence of COVID-19 in Raparin July 2020 Vol. 23 Issue 11

and respond to various infection, and also it was found that at elderly people consecutive BCG vaccine reduced upper respiratory tract infections [25-28] therefore BCG might have a role in protecting Raparin district people against severe coronavirus disease 2019. In conclusion, it can be demonstrated thatthere are many factors that might less coronavirus prevalence. The factors are quarantining, BCG vaccination, sharing and practicing health guidance, preventing social gathering, living in a house rather than flat, the lockdown of shopping, university and all schools and administrations, prevention transportation and the most important was the establishment a center for combating coronavirus before spreading and having any positive cases in Kurdistan.

ACKNOWLEDGMENT We owe special appreciation togeneraldirectorate of health-Raparinfor giving us the opportunity of joining this humanitarian and ethical work.The deepest gratitude goes to;IsmahilHamadaminHamad, Rizgar Mustafa Ismahil and Aras MuhamadHamad for their support and cooperation during the laboratory work. REFERENCES 1- Callaway, E., 2020. The race for coronavirus vaccines: a graphical guide. Nature, 580(7805), p.576.

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