【病毒外文文献】2004 Detection of Feline Coronavirus Antibody, Feline Immunodeficiency Virus Antibody, and Feline Leukemia Virus Antigen
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NOTE Internal Medicine Detection of Feline Coronavirus Antibody Feline Immunodeficiency Virus Antibody and Feline Leukemia Virus Antigen in Ascites from Cats with Effusive Feline Infectious Peritonitis Takehisa SOMA 1 and Hiroshi ISHII 2 1 Marupi Lifetech Co Ltd 103 Fushiocho Ikeda Osaka 563 0011 and 2 Sukagawa Animal Hospital 44 11 Morijuku Sukagawa Fukushima 962 0001 Japan Received 19 June 2003 Accepted 20 August 2003 ABSTRACT To investigate the usefulness of ascites as a material for viral tests in cats with effusive feline infectious peritonitis FIP we attempted to detect anti feline coronavirus antibody anti feline immunodeficiency virus antibody and feline leukemia virus antigen in ascites from 88 cats clinically suspected with effusive FIP In each of these three viral tests all cats positive for serum an tibody antigen were also positive for ascitic antibody antigen while cats negative for serum antibody antigen were also negative for ascitic antibody antigen This finding indicates that ascites is useful for these viral tests KEY WORDS ascites FIP viral test J Vet Med Sci 66 1 89 90 2004 Feline infectious peritonitis FIP is a progressive viral infectious disease caused by feline coronavirus FCoV infection in domestic and wild large cats 1 As a diagnos tic method of FIP anti FCoV antibody test has been reported in 1976 8 and this test is now used as an impor tant diagnostic method of FIP Superinfection of FCoV infected cats with feline leukemia virus FeLV or feline immnunodeficiency virus FIV has been reported as a fac tor inducing FIP 3 9 and tests of these two viruses in addition to anti FCoV antibody test are important for the diagnosis of FIP FIP is classified into the effusive and non effusive forms The former is characterized by accumula tion of ascites and thoracic effusion and the latter is characterized by multiple pyogranulomous lesions Gener ally symptoms of the former acutely progress compared to the latter 1 and a sufficient amount of serum cannot be collected due to emaciation in many cases of the effusive FIP In cases of the effusive FIP immune complexes injure the blood vessel wall and cause leakage of plasma proteins and body fluid from the blood vessel into the body cavities 5 Accordingly certain amounts of serum viral antigens and antibodies may transfer to exudates However there has been no report of comparison of viral tests in serum and exudates In this study to investigate the usefulness of peritoneal exudates ascites as a material for viral tests in cats with effusive FIP we attempted to detect anti FCoV antibody anti FIV antibody and FeLV antigen in ascites from cats clinically suspected with effusive FIP and compared with the serum levels As test materials blood and ascites were collected from 88 pet cats clinically suspected of having FIP due to accu mulation of ascites and the supernatants after centrifugation at 3 000 rpm for 15 min were tested Anti FCoV antibody test was performed by indirect fluorescent antibody IFA assay as reported by Pedersen et al 7 Confluent mono layers of Crandell feline kidney CRFK cells American Type Culture Collection ATCC U S A were infected with FIP virus 79 1146 strain ATCC U S A in a 96 well tissue culture plate After 20 hr incubation the plate was fixed with methanol acetone solution 1 3 The sample that was diluted 1 100 and then serially 2 fold was added into the plate After incubation the plate was washed and fluores cein isothiocyanate FITC conjugated anti cat IgG goat antibody Cappel U S A was added into each well After incubation the plate was washed and examined with a fluo rescent microscope The antibody titer was expressed as the highest sample dilution with the positive reaction Anti FIV antibody test was performed by IFA assay as reported by Barr et al 2 FIV chronically infected CRFK cells ATCC U S A were grown in a 96 well tissue culture plate The plate was fixed with the methanol acetone solu tion The sample that was diluted 1 50 and then serially 2 fold was added into the plate After incubation the plate was washed and the FITC conjugated anti cat IgG goat anti body was added into each well After incubation the plate was washed and examined with the fluorescent microscope The antibody titer was expressed as the highest sample dilu tion with the positive reaction FeLV antigen test was per formed by enzyme linked immunosorbent assay ELISA as reported by Lutz et al 6 The sample diluted 1 8 was added into the 96 well ELISA plate coated with anti p27 mouse monoclonal antibody ATCC U S A After incu bation the plate was washed and horseradish peroxidase conjugated anti p27 goat antibody ViroStat U S A was added into each well After incubation the plate was washed and the substrate buffer 0 05 M citric acid contain ing 0 2 mM 2 2 azino bis 3 ethylbenzothiazoline 6 sul fonic acid and 0 004 H 2 O 2 pH 4 0 was added into each well After incubation optical density OD value was determined at 405 nm When an anti FCoV antibody titer of 1 100 or more wasT SOMA AND H ISHII 90 regarded as positive all 62 cats positive for serum antibody were also positive for ascitic antibody while 26 cats nega tive for serum antibody were also negative for ascitic anti body Table 1 Next the serum and ascitic antibody titers were compared in the positive cats The ascitic antibody titer was the same to the serum antibody titer in 48 cats The ascitic antibody titers were 1 2 and 1 4 of the serum anti body titer in 12 and two cats respectively Table 2 When an anti FIV antibody titer of 1 50 or more was regarded as positive all 10 cats positive for serum antibody were also positive for ascitic antibody while 78 cats negative for serum antibody were also negative for ascitic antibody Table 1 The ascitic antibody titer was the same to the serum antibody titer in 7 cats The ascitic antibody titers were 1 2 and 1 4 of the serum antibody titer in two and one cats respectively Table 2 When an FeLV antigen titer OD value of 0 100 or more was regarded as positive all 16 cats positive for serum antigen were also positive for ascitic antigen while 72 cats negative for serum antigen were also negative for ascitic antigen Table 1 The mean and stan dard deviation of the ascitic OD value serum OD value ratio were 1 45 0 49 in the positive cats As described above all cats positive for serum antibody antigen were also positive for ascitic antibody antigen while cats negative for serum antibody antigen were also negative for ascitic antibody antigen in each of these three viral tests This finding indicates that ascites is useful for the viral tests and the viral tests using ascites can be per formed in cats in which blood sampling is difficult It has been reported that the IgG antibody level is slightly lower about 70 in exudates than in serum in humans 10 The anti FCoV and anti FIV antibody titers were lower 1 2 1 4 in the ascites than in the serum in 22 6 and 30 0 of the antibody positive cats respectively in the antibody tests that are IgG antibody measurement systems in this study In contrast the FeLV antigen titer in the ascites was 1 45 fold higher on average than that in the serum in the antigen positive cats It has been reported that the ratio of exudates serum proteins was inversely related to the molecular weight in humans 11 The FeLV antigen test performed in this study detects p27 viral core antigen with a molecular weight of 27 k Dalton 6 The leakage rate of this antigen may have been higher than that of IgG antibody with a molecular weight of 150 k Dalton 4 It will be necessary to conduct further studies using a larger sample for a reliable comparison of the serum and ascitic titers in cats with FIP Exudates are generally more viscous than serum in FIP 12 In this study some ascites samples were very viscous but the test systems of this study could readily measure the titers In Japan simple viral diagnostic kits have been com monly used as rapid tests in animal hospitals in recent years In some kits the test material is dripped on the kit without dilution and accurate results might not be obtained from viscous materials To increase the usefulness of exudates it is necessary to investigate applicability of exudates to such diagnostic kits REFERENCES 1 Addie D D and Jarrett O 1998 pp 58 69 In Infectious Dis eases of the Dog and Cat Greene C E ed WB Saunders Company Philadelphia 2 Barr M C Dough M B Jacobson R H and Scott F W 1991 J Am Vet Med Assoc 199 1377 1381 3 Cotter S M Gilmore C E and Rollins C 1973 J Am Vet Med Assoc 162 1054 1058 4 Greene C E 1984 pp 35 66 In Clinical Microbiology and Infectious Diseases of the Dog and Cat Greene C E ed WB Saunders Company Philadelphia 5 Grahn B H 1991 Vet Med 86 376 393 6 Lutz H Pedersen N C Durbin R and Theilen G H 1983 J Immunol Method 56 209 220 7 Pedersen N C Boyle J F and Floyd K 1981 Am J Vet Res 42 363 367 8 Pedersen N C 1976 Am J Vet Res 37 1449 1453 9 Poland A M Vennema H Foley J E and Pedersen N C 1996 J Clin Microbiol 34 3180 3184 10 Rerabek J E 1977 Clin Chim Acta 76 363 369 11 Telvi L Jaubert F Eyquem A Andreux J P Labrousse F and Chretien J 1979 Thorax 34 389 392 12 Weiss R C 1991 Vet Med 86 308 319 Table 2 Ratio of antibody titers in the ascites and serum from the antibody positive cats Ascites Serum a FCoV antibody FIV antibody 1 48 77 4 7 70 0 1 2 12 19 4 2 20 0 1 4 2 3 2 1 10 0 Total 62 100 10 100 a Ascites Serum Reciprocal of antibody titer in the ascites reciprocal of antibody titer in the serum Table 1 Comparison of viral test results in the ascites and serum from 88 cats suspected of having FIP Serum FCoV antibody 6 20 02 6 Ascites FIV antibody 1 00 07 8 FeLV antigen 1 60 07 2 Positive Negative An anti FCoV antibody titer of 1 100 or more an anti FIV antibody titer of 1 50 or more and an FeLV antigen titer of 0 100 or more were regarded as positive- 配套讲稿:
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