The virus strains dromedary MERS-CoV Al-Hasa KFU-HKU13 2013 (Al-Hasa 13) and dromedary MERS-CoV Egypt NRCE-HKU270 2013 (Egypt 270) were isolated in our laboratory as previously described [10,12]

The virus strains dromedary MERS-CoV Al-Hasa KFU-HKU13 2013 (Al-Hasa 13) and dromedary MERS-CoV Egypt NRCE-HKU270 2013 (Egypt 270) were isolated in our laboratory as previously described [10,12]. has been used to minimise the human health risk from highly pathogenic avian influenza H5N1 and SARS [3,4]. Sero-epidemiology is an invaluable tool in such investigations. Many sero-epidemiological studies on domestic livestock have reported high sero-prevalence in dromedary camels in the Arabian peninsula and Africa [5C8]. The detection of MERS-CoV computer virus by MSI-1701 reverse transcription polymerase chain reaction (RT-PCR) and computer virus isolation supports these sero-epidemiological findings and the contention that dromedaries are a natural host for MERS-CoV [9C11]. But it is not clear if MSI-1701 dromedaries are the main source of human infection. We had previously reported a MERS-CoV pseudoparticle neutralization assay (ppNT) that can be used to detect antibody to MERS-CoV without the need for Biosafety Level-3 (BSL-3) containment that is required for conventional MERS-CoV microneutralisation (MN) assessments [6]. In this study, we systematically investigate potential cross reactions that may confound the use of these two MSI-1701 assays in sero-epidemiological studies in animals. Methods Viruses MERS-CoV EMC strain was provided TIAM1 by Dr Ron Fouchier, Erasmus Medical Centre, Rotterdam. The computer virus strains dromedary MERS-CoV Al-Hasa KFU-HKU13 2013 (Al-Hasa 13) and dromedary MERS-CoV Egypt NRCE-HKU270 2013 (Egypt 270) were isolated in our laboratory as previously described [10,12]. The viruses were cultured and titrated in Vero cells (ATCC CCL-81). Sera Immune sera specific for alphacoronaviruses (porcine respiratory coronavirus, feline infectious peritonitis computer virus, canine coronavirus and porcine transmissible gastroenteritis computer virus), betacoronaviruses (mouse hepatitis computer virus: strains JHM and A59, SARS coronavirus, BCoV) and gammacoronavirus (infectious bronchitis computer virus) were obtained from BEI-Resources (animal CoV reagents supplied to BEI by Dr Linda Saif) (http://www.beiresources.org/About/BEIResources.aspx) or generated by Dr Linda Saif or Dr Stanley Perlman, as indicated in table 1. The homologous antibody titres to the immunising computer virus was also obtained from the respective sources supplying these antisera (Table 1). Table 1 Cross neutralization antibody titers for MERS-CoV and BCoV in anti-sera raised against different coronaviruses

Genus Antisera Homologous Ab titre by ELISA unless otherwise specified MERS-CoV MN titre MERS-CoV ppNT titre BCoV MN titre

Alpha coronavirusGnotobiotic pig antiserum to Porcine Respiratory coronavirus – NR MSI-1701 4601:1200#<1:10<1:10<1:10Guinea Pig antiserum to Feline infectious Peritonitis computer virus - NR 25181:2000#<1:10<1:10<1:10Guinea pig antiserum to Canine Coronavirus - NR 27271:4094?<1:10<1:10<1:10Gnotobiotic pig antiserum to Porcine transmissible gastroenteritis virus - NR 4581:1400#<1:10<1:10<1:10Beta coronavirusGuinea Pig anti-SARS-CoV - NR 103611:2560<1:10<1:10<1:10Rabbit anti serum for SARS-CoV S protein (Zero titer) - NRC 769<1:10<1:10<1:10<1:10Rabbit anti serum for SARS-CoV S protein (low titer) - NRC 7701:80<1:10<1:10<1:10Rabbit anti serum for SARS-CoV S protein (medium titer) - NRC 7711:160<1:10<1:10<1:10Rabbit anti serum for SARS-CoV S protein (high titer) - NRC 7721:640<1:10<1:10<1:10Mouse hepatitis virus (JHM strain) hyper immunized mouse dam 11:1778?Neutralization titre<1:10<1:10<1:10Mouse hepatitis computer virus (JHM strain) hyper- immunised mouse dam 21:363?Neutralization titre<1:10<1:10<1:10Mouse hepatitis computer virus (A59 strain) infected mouse1:1000?Neutralization titre<1:10<1:10<1:10BCoV antisera from guinea pig - GP 99101:20480?<1:10<1:101:160BCoV antisera from germfree calf - NR 4561:10000#<1:10<1:101:40BCoV antisera from germfree calf - C38161:580?Neutralization titre<1:10<1:101:640Gamma coronavirusGuinea Pig anti serum to infectious bronchitis computer virus - NR 25151:50000#<1:10<1:10<1:10 Open in a separate windows Homologous antibody titre data from #BEI resources; ?Linda Saif; ?Perlman Stanly. All the antibody titres are acquired within this scholarly research. All homologous antibody titres MSI-1701 are titres aside from antisera ELISA.