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1、RESEARCH ARTICLE Countrywide Survey for MERS-Coronavirus Antibodies in Dromedaries and Humans in Pakistan Ali Zohaib1,2Muhammad Saqib3Muhammad Ammar Athar4Jing Chen1Awais-ur-Rahman Sial5 Saeed Khan6Zeeshan Taj7Halima Sadia8Usman Tahir8Muhammad Haleem Tayyab3 Muhammad Asif Qureshi6Muhammad Khalid Man
2、soor9Muhammad Ahsan Naeem10Bing-Jie Hu1 Bilal Ahmed Khan6Ikram Din Ujjan11Bei Li1Wei Zhang1Yun Luo1,2Yan Zhu1Cecilia Waruhiu1,2 Iahtasham Khan12Xing-Lou Yang1Muhammad Sohail Sajid3Victor Max Corman13,14Bing Yan1 Zheng-Li Shi1 Received: 26 July 2018/Accepted: 3 September 2018 ? Wuhan Institute of Vir
3、ology, CAS and Springer Nature Singapore Pte Ltd. 2018 Abstract Middle East Respiratory Syndrome Coronavirus (MERS-CoV) is a zoonotic pathogen capable of causing severe respiratory disease in humans. Although dromedary camels are considered as a major reservoir host, the MERS-CoV infection dynamics
4、in camels are not fully understood. Through surveillance in Pakistan, nasal (n = 776) and serum (n = 1050) samples were collected from camels between November 2015 and February 2018. Samples were collected from animal markets, free-roaming herds and abattoirs. An in-house ELISA was developed to dete
5、ct IgG against MERS-CoV. A total of 794 camels were found seropositive for MERS-CoV. Prevalence increased with the age and the highest seroprevalence was recorded in camels aged10 years (81.37%) followed by those aged 3.110 years (78.65%) and B 3 years (58.19%). Higher prevalence was observed in fem
6、ale (78.13%) as compared to male (70.70%). Of the camel nasal swabs, 22 were found to be positive by RT-qPCR though with high Ct values. Moreover, 2,409 human serum samples were also collected from four provinces of Pakistan during 20162017. Among the sampled population, 840 humans were camel herder
7、s. Although we found a high rate of MERS-CoV antibody positive dromedaries (75.62%) in Pakistan, no neutralizing antibodies were detected in humans with and without contact to camels. Keywords Middle East Respiratory Syndrome Coronavirus (MERS-CoV) ? Camel ? Human ? Pakistan Gossner et al. 2016; Haa
8、gmanset al. 2014; Alraddadi et al. 2016). Moreover, large hospital linked outbreaks have also been reported from cases imported to other countries (Park et al. 2017). Accumulating evidence sug- gests that camels are reservoir for MERS-CoV. Although camel to human transmission has been documented, mo
9、st of human infections are due to human to human trans- mission especially in healthcare settings (Park et al. 2017). In a geographically comprehensive study from KSA, MERS-CoV antibodies were detected in approximately 0.15% of the sampled human population, depicting spo- radic infections without se
10、vere disease (Mu lleret al. 2015). A signifi cantly higher seroprevalence was observed in slaughterhouse workers and camel shepherds (Mu ller et al. 2015). High prevalence of MERS-CoV antibodies has been reported in camels from different countries of Arabian Peninsula (Saudi-Arabia, United Arab Emir
11、ates, Qatar, and Oman) and of Africa (Kenya, Sudan, Nigeria, Burkina Faso, Ethiopia and Morocco) (Gossneret al. 2016; Werneryet al. 2015; Haagmanset al. 2014; Reusken et al. 2013; Munyua et al. 2017; Mu ller et al. 2014; Chu et al. 2018). Presence of MERS-CoV neutralizing anti- bodies in archived ca
12、mel sera from 1983 suggests long- term circulation of virus among camel population (Mu ller et al. 2014). However, recent studies from Australia, China and Kazakhstan suggest no evidence of MERS-CoV infection in dromedary and Bactrian camels (Crameri et al. 2015; Liu et al. 2015; Miguel et al. 2016)
13、. A small-scale study from Punjab, Pakistan has reported a high percentage of up to 39.5% of dromedaries having neutralizing antibodies against MERS-CoV (Saqib et al. 2017). This is contrasted by the lack of reported human MERS-CoV infection in Pakistan. In view of uncertain disease prevalence, limi
14、ted capacity for routine surveil- lance and considerably large human and dromedary pop- ulation, conducting a countrywide cross-sectional study for MERS-CoV infections in dromedaries and human is of interest for global public health agencies. The aim of this study was to determine the country-wide p
15、revalence of MERS-CoV in camel and human population of Pakistan. Materials and Methods Study Locales and Sampling This cross-sectional study was designed to determine the prevalence of MERS-CoV in camel and human population of Pakistan. For camel sampling, sites were chosen based on dromedary popula
16、tions and the presence of veterinary clinics (Fig. 1). During 20152018, 776 camel nasal swabs were collected. After collection, swabs were placed into tubes containing RNAlater?(Ambion, Austin, USA) and stored at - 80 ?C. A total of 1050 dromedary sera were also collected using blood collection system (Becton Dickinson Co, San Jose, USA). Before taking blood sam- ples the clinical parameters including rectal temperature, body condition and symptoms of any disease were also recorded. For human s
