Abstract
A reversed passive latex agglutination test (RPLA) using anti-canine haemoglobin (Hb) antibody was developed for detecting bleeding in the lower digestive organs in dogs, and its applicability as a simple test for faecal occult blood was assessed. In Ouchterlony's gel immunodiffusion test, the anti-canine Hb antibody used to sensitize the latex reacted with canine Hb but not with Hbs, plasmas or meat extracts from pigs, goats, sheep, cattle, horses or chickens, or with fish extracts. Using latex sensitized with 50 µg/mg of anti-canine Hb IgG antibody, the lowest limit of detection for canine Hb was 21 µg/ml, and the latex reacted negatively with all test specimens other than canine Hb. In an in vitro experiment with a mixture of canine faeces and erythrocytes, the antigenicity of the Hb was found to undergo only very slight changes even when the specimens were allowed to stand for 12 h at room temperature. Hb could not be detected by RPLA in any of four successive faecal samples from three experimental dogs after infusion of autologous blood (5, 3 or 1 ml) into the stomach. In 3 other experimental dogs given an infusion of autologous blood (5, 3 or 1 ml) into the ascending colon, the presence of Hb was confirmed by RPLA in all four successive faecal samples obtained from those which received 5 or 3 ml of blood and in all except that obtained following the first defecation from the animal which had received 1 ml of blood.
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REFERENCES
Adams, E.C. and Layman, K.M., 1974. Immunochemical confirmation of gastrointestinal bleeding. Annals of Clinical Laboratory Science, 4, 343-349
Ahlquist, D.A., McGill, D.B., Schwartz, S., Taylor, W.F., Ellefson, M. and Owen, R.A., 1984. Hemo-Quant, a new quantitative assay for fecal hemoglobin: comparison with Hemoccult. Annals of Internal Medicine, 101, 297-302
Ahlquist, D.A., McGill, D.B., Schwartz, S., Taylor, W.F. and Owen, R.A., 1985. Fecal blood levels in health and disease: a study using Hemo-Quant. New England Journal of Medicine, 312, 1422-1428
Barrows, G.H., Burton, R.M., Jarrett, D.D., Russell, G.G., Alford, M.D. and Songster, C.L., 1978. Immunochemical detection of human blood in feces. American Journal of Clinical Pathology, 69, 342-345
Bradford, M.M., 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248-254
Burton, R.M., Landreth, K.M., Barrows, G.H., Jarrett, D.D. and Songster, C.L., 1976. Appearance, properties, and origin of altered human hemoglobin in feces. Laboratory Investigation, 35, 111-115
Francis, R.T. Jr, Booth, J.W. and Becker, R.R., 1985. Uptake of iron from hemoglobin and the haptoglobin-hemoglobin complex by hemolytic bacteria. International Journal of Biochemistry, 17, 767-773
Kachmar, J.F., 1970. Tests for occult blood. In: N.W. Tietz (ed.), Fundamentals of Clinical Chemistry, (W.B. Saunders, Philadelphia), 279-282
Macrae, F.A. and St John, D.J.B., 1982. Relationship between patterns of bleeding and hemoccult sensitivity in patients with colorectal cancers or adenomas. Gastroenterology, 82, 891-898
Oshima, M., Watanabe, K., Sasaki, T., Fujimasa, A., Takeda, J., Kakegawa, T. and Hara, M., 1985. ELISA method for the detection of occult blood in feces. Journal of Clinical and Experimental Medicine, 135, 261-263 (in Japanese)
Ostrow, J.D., Mulvaney, C.A., Hansell, J.R. and Rhodes, R.S., 1973. Sensitivity and reproducibility of chemical tests for fecal occult blood, with an emphasis on false-positive reactions. American Journal of Digestive Diseases, 18, 930-940
Peterson, W.L. and Fordtran, J.S., 1985. Quantitating the occult. New England Journal of Medicine, 312, 1448-1450
Saito, H., Tsuchida, S., Kakizaki, R., Fukushi, M., Sano, M., Aizawa, N., Munakata, A. and Yoshida, A., 1984. An immunological fecal occult blood test for mass screening of colorectal cancer by reversed passive hemagglutination (RPHA). Japanese Journal of Gastroenterology, 81 2831 (in Japanese)
Schwartz, S., Dahl, J., Ellefson, M. and Ahlquist, D., 1983. The “Hemo-Quant” test: a specific and quantitative determination of heme (hemoglobin) in feces and other materials. Clinical Chemistry, 29, 2061-2067
Simon, J.B., 1985. Occult blood screening for colorectal carcinoma: a critical review. Gastroenterology, 88, 820-837
Takeshita, T., Horiguchi, J., Kinoshita, T., Kubota, Y., Chen, P.-C., Katsumata, S., Horimukai, F., Miyaoka, M., Matsumoto, E., Saito, T. and Ashizawa, S., 1985. Latex agglutination test for fecal occult blood. Journal of the Japan Society of Colo-Proctology, 38, 780-783 (in Japanese with English summary)
Towbin, H., Staehelin, T. and Gordon, J., 1979. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proceedings of the National Academy of Sciences of the USA, 76, 4350-4354
Turunen, M.J., Liewendahl, K., Partanen, P. and Adlercreutz, H., 1984. Immunological detection of faecal occult blood in colorectal cancer. British Journal of Cancer, 49, 141-148
Uchida, K., Matsuse, R., Miyachi, N. and Tomita, S., 1990. Immunological fecal occult blood test. Japanese Journal of Clinical Medicine (Supplement), 48, 1276-1285 (in Japanese)
Yamamoto, S., Tagata, K., Ishikawa, Y., Fujise, H., Nagahata, H., Yamada, M., Sakano, T., Morimatsu, M. and Naiki, M., 1992. Preparation of latex sensitized with rabbit IgG antibody for slide reversed passive agglutination.Veterinary Research Communications, 16, 265-272
Yamamoto, S., Abe, N., Santsuka, H., Shida, T., Kishida, K., Kuwajima, S., Yamada, M., Morimatsu, M. and Naiki, M., 1993. Efficient preparation of monospecific anti-canine C-reactive protein serum and purification of canine C-reactive protein by affinity chromatography. Veterinary Immunology and Immunopathology, 36, 293-301
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Jinbo, T., Shinmura, R., Shida, T. et al. Experimental Detection of Canine Haemoglobin (Occult Blood) in Canine Faeces by Reversed Passive Latex Agglutination. Vet Res Commun 21, 347–353 (1997). https://doi.org/10.1023/A:1005812304532
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DOI: https://doi.org/10.1023/A:1005812304532