The enzyme-linked immunosorbent assay using the so called "double-sandwich" tecnique was applied to determine C. boturinum type E and F toxins simultaneously. Polystyrene tubes were coated with horse anti-type E and F toxins serum and then sample was added. The tubes were subsequently treated with rabbit anti-type E and F toxins IgG, and sheep anti-rabbit serum IgG-horseradish peroxidase conjugate.
The minimum detectable dose of type E and F toxins in culture filtrate and in food sample was 10 mouse ip LD50.
The cross-reaction value of type E toxins against type F anti-serum was 0.062 and that of type F toxin against type E anti-serum was 0.081.
Clostridium botulinum type E, strain Alaska 와 type F, strain Lageland의 독소를 간단히, 동시에 검출할 수 있는 면역학적 효소 방법을 연구하였다. 방법은 다음과 같다.
(1) Horse anti-type E and type F serum 을 polystyrene tube 에 부착
(2) 각기 다른 양의 type E, type F, type E+type F 독소를 가함
(3) PTB 로 희석시킨 rabbit anti-type E and type F IgG 를 가함.
(4) PTB 로 희석시킨 sheep anti-rabbit IgG-HRPO cojugate 를 가함.
(5) Substrate 가함.
(6) 반응을 정지시키고 Abs.(449nm)record. 한 tube 내에서 type Eand F 독소 검출이 가능하며, 검출 한계는 약 10mouse ip $LD_50$ 였으며, 또한 음식 시료에서의 독소 검출도 가능함을 볼 수 있다.
