Development of a canine trypsin-like immunoreactivity assay system using monoclonal antibodies

Waritani, Takaki; Okuno, Yoko; Ashida, Yoshinori; Hisasue, Masaharu; Tsuchiya, Ryo; Kobayashi, Kosaku; Yamada, Takatsugu

Development of a canine trypsin-like immunoreactivity assay system using monoclonal antibodies

2002

The radioimmunoassay (RIA) for trypsin-like immunoreactivity (TLI) is one of the most sensitive and specific tests for detecting exocrine pancreatic insufficiency (EPI). An abnormally low serum TLI concentration (<2.5 ng/ml) indicates end-stage EPI. Although RIA methods can be used to detect canine serum TLI, these procedures are beyond the capabilities of most veterinary clinics and general laboratories. Using monoclonal antibodies (mAbs), we developed an enzyme-linked immunosorbent assay (ELISA) for canine TLI and incorporated it into an immunochromatographic test (ICT) for the diagnosis of EPI. The ELISA was linear over TLI concentrations of 1–100 ng/ml. Levels of intra-assay coefficients of variance (CVs) were 1.8–6.1%, inter-assay CVs were 5.1–9.8%, and the recovery of TLI added to two samples of canine serum ranged from 89 to 111 and 93 to 108%, respectively. Good correlation (correlation coefficient, 0.974) occurred between the TLI values obtained by the ELISA method and those by RIA from 56 clinical samples. Serum TLI values in clinically healthy dogs ranged from 7.8 to 29.2 ng/ml by ELISA, and those from dogs with EPI were 0.0–0.6 ng/ml. The values were 0.0–287.4 ng/ml for dogs with pancreatitis, and those from dogs with gastrointestinal disease were 5.5–58.9 ng/ml. The only statistically significant difference (P<0.01) occurred between the TLI level of healthy dogs and those with EPI. The ICT kit showed high reproducibility, and the TLI values yielding negative results differed significantly (P<0.01) from those returning positive results. The ICT kit yielded negative results (indicating EPI) from clinical serum samples with TLI concentrations of 0.0–4.1 ng/ml by ELISA. Both the ELISA and ICT kit are useful tools in the diagnosis of canine EPI.

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