Sharif et al. Acta Veterinaria Scandinavica 2010, 52:1 http://www.actavetscand.com/content/52/1/1 Page 2 of 7 Figure 1 Distribution of feline coronavirus positive cats categorized by age, breed and gender. DSH: Domestic Short Hair retrospective study [11]. Antibodies against FCoVs were found in 100% of cats living in Malaysian catteries [12] and the virus was detected in 84% of healthy cats using RT-PCR [13]. In present study, a conserved region of 3’untranslated region (3’UTR) is used to detect FCoV and determine the descriptive distribution and phylo- geny of local isolates in FIP-suspected cats. Abdominal fluids and/or tissue samples of 28 cats sus- pected of having the effusive form of FIP were obtained from the University Veterinary Hospital, Universiti Putra Malaysia (UVH-UPM) over the period of three years (2007-2009). Ascitic fluids were diluted 1:10 in phosphate buffer solution (PBS), aliquoted and stored at -70°C until used. Organ samples were homogenized in 1:10 of PBS. Insoluble components were removed by centrifugation for 10 min at 3000 g and the supernatant fraction was collected and kept at -70°C. Two FCoV reference strains (FECV 79-1683; ATCC® No.VR-989™ and FIPV79-1146; ATCC® No. VR-216™) were used for RT-PCR optimization. Virus stocks were propagated in confluent Crandell Feline Kidney cells. The viruses were harvested when the infected cells showed 80% cyto- pathic effects. The virus suspension was freezed-thawed three times and stored at -70°C until used. RNA was extracted from the infected cell culture supernatants and clinical samples using TRIZOL® Reagent (Invitrogen, Carlsbad, California, USA) accord- ing to the manufacturer’s instructions. The partial 3’UTR was amplified by RT-PCR using previously described primers [7]. One-step RT-PCR was performed using Access RT-PCR System and RNasin® Ribonuclease Inhibitor (Promega, Madison, Wisconsin, USA). The Table 1 Statistical analysis of feline infectious peritonitis suspected cats tested for feline coronavirus (FCoV) by RT-PCR assay. Age Breed Gender Criteria < 2 years ≥ 2 years DSH Persian Siamese Male Female No. of tested cats 17 11 17 10 1 21 7 No. of FCoV-positive cats 16 9 14 10 1 20 5 FCoV-positivity (%) 94 82 82 100 100 95 71 Odds 16 4.5 4.67 * * 20 2.5 DSH: Domestic Short Hair * Insufficient number of cats to allow statistical calculations. Odds Ratio 3.5556 Confidence Interval 0.2816 to 44.886 * * * 8 * * * 0.5985 to 106.9411