Sharif et al. Acta Veterinaria Scandinavica 2010, 52:1 http://www.actavetscand.com/content/52/1/1 Table 2 List of feline coronavirus isolates and strains included in the sequence and phylogenetic analysis. No. Isolate/Strain Accession No. Origin Reference 1 UPM1C/07 FJ897745 Malaysia This paper 2 UPM2C/07 FJ897746 Malaysia This paper 3 UPM3C/07 FJ897747 Malaysia This paper 4 UPM4C/08 FJ897748 Malaysia This paper 5 UPM5C/08 FJ897749 Malaysia This paper 6 UPM6C/08 FJ897750 Malaysia This paper 7 UPM7C/09 FJ897751 Malaysia This paper 8 UPM8C/09 FJ897752 Malaysia This paper 9 UPM9C/09 FJ897753 Malaysia This paper 10 UPM10C/09 FJ897754 Malaysia This paper 11 UPM28C/08 GQ233036 Malaysia [19] 12 UPM29C/08 GQ233037 Malaysia [19] 13 UPM30C/09 GQ233038 Malaysia [19] 14 UPM31C/09 GQ233039 Malaysia [19] 15 UU10 FJ938059 Netherlands Unpublished 16 UU15 FJ938057 Netherlands Unpublished 17 UU11 FJ938052 Netherlands Unpublished 18 UU9 FJ938062 Netherlands Unpublished 19 UU3 FJ938061 USA Unpublished 20 UU2 FJ938060 USA Unpublished 21 RM FJ938051 USA Unpublished 22 UCD11b-2b FJ917535 USA Unpublished 23 UCD11b-2a FJ917534 USA Unpublished 24 UCD11b-1b FJ917533 USA Unpublished 25 UCD11b-1a FJ917532 USA Unpublished 26 UCD11a-1b FJ917531 USA Unpublished 27 UCD11a-1a FJ917530 USA Unpublished 28 UCD17 FJ917527 USA Unpublished 29 UCD14 FJ917524 USA Unpublished 30 UCD13 FJ917523 USA Unpublished 31 UCD5 FJ917522 USA Unpublished 32 UCD12 FJ917521 USA Unpublished 33 UCD11b FJ917520 USA Unpublished 34 UCD11a FJ917519 USA Unpublished 35 Black EU186072 USA [3] 36 NTU2/R/2003 DQ160294 Taiwan Unpublished 37 UU16 FJ938058 Netherlands Unpublished 38 UU5 FJ938056 Netherlands Unpublished 39 UU8 FJ938055 Netherlands Unpublished 40 UU7 FJ938053 Netherlands Unpublished 41 UCD18b FJ917529 USA Unpublished 42 UCD18a FJ917528 USA Unpublished 43 UCD16 FJ917526 USA Unpublished 44 UCD15a FJ917525 USA Unpublished 45 DF-2 DQ286389 USA Unpublished 46 C1Je DQ848678 UK Unpublished 47 NTU156/P/2007 GQ152141 Taiwan Unpublished 48 UU4 FJ938054 Netherlands Unpublished 49 79-1146 DQ010921 USA [21] 50 Wellcome X90571 Netherlands [22] Page 3 of 7 Table 2: List of feline coronavirus isolates and strains included in the sequence and phylogenetic analysis. (Continued) 51 52 53 54 55 56 57 58 59 60 61 62 UCD1 UCD TN406 UCD3a UCD2 Dahlberg UCD3 UCD4 NOR15 UCD12-1 UCD6-1 79-1683 X90575 X90574 X90570 FJ943761 X90576 X90572 X90577 X90578 X90573 FJ943766 FJ943772 X66718 USA [22] USA [22] Netherlands [22] USA Unpublished USA [22] Netherlands [22] USA [22] USA [22] Netherlands [22] USA Unpublished USA Unpublished USA [23] reaction was optimized on a thermal cycler (MJ Research, Waltham, Massachusetts, USA). PCR products of 223 bp were analyzed using electrophoresis on a 2% agarose gel, stained with ethidium bromide and observed under UV light. PCR products of 10 positive cases were selected randomly, purified using PCR SV protocol (GENEALL®, Seoul, South Korea) and sequenced in both direction with the primers (Medi- gene, Selangor, Malaysia). Data analysis was performed using Statistical Tables Calculator, which is available online at http://faculty.vas- sar.edu/lowry/odds2x2.html. Age, breed and gender dif- ferences were compared by calculating positivity rate, odds and 95% confidence intervals. The RT-PCR assay amplified the target band in 25 out of 28 cats’ samples (89%). Although, the PCR results must be interpreted in conjunction with clinical or pathological findings, detection of the virus in FIP-sus- pected cats may be useful to confirm FIP. Since FCoVs are ubiquitous in cats with high seroprevalence [5,6,12], PCR provides the obvious advantage over serology by directly detecting FCoV genome rather than document- ing a previous immune system encounter with the coro- navirus. The primers of this PCR assay were chosen from a highly conserved region of 3’UTR of the FCoV genome to detect most, if not all of the FCoV strains. The usefulness of these primers for a general screening test has been reported previously [14-16]. FCoV-positivity rate in cats younger than two years old (64%) was higher than older cats, but they are not significant. However, the result is consistent with other studies demonstrating higher incidence of FIP in cats below 2 years of age [5,11,14] and agree with the fact that FIP is a disease of young cats. Typical clinical cases are first appear during the postweaning period, but most deaths from FIP occur in cats 3-16 months of age [6]. Most of the FCoV-positive cats in our study were males (80%). Higher incidence of FIP among males was