Study of serological and genetical diagnoses for B virus infection in monkeys

1.用酶免疫法对日本国立大学实验动物协会实验动物设施饲养的962份健康非人灵长类动物血清进行了B型疱疹病毒(脑疱疹病毒1型)的血清学调查。抗体阳性率为40%(384/961)。猕猴的抗体效价(60%的食蟹猴、53%的恒河猴和34%的日本猴)高于非猕猴物种(21%)。这些数据也对日本猴子在生命科学研究领域的使用发出了警告。我们研究了用替代或替代抗原代替危险的B病毒：1)人类疱疹病毒1型(HSV1)，2)疱疹病毒乳头状病毒2型(HVP2)抗原。HSV1(gP和gD抗原)与B病毒EL ISA的符合率为96%。另一方面，用来自美国和日本的350份猴血清标本进行检测，其与B病毒EL ISA的符合率为98%。这些数据提示可采用替代方法进行抗体检测，尤其是对HvP2抗原的检测。通过尝试RT-巢式聚合酶链式反应方法，在日本来源的抗体阳性的21只日本猴的三叉神经节标本中，有3份样本可以检测到B病毒基因组。它是利用BV的两种糖蛋白抗原作为重组抗原而实现的。然而，他们的产品对哺乳动物细胞具有细胞毒性作用，因此被判定为不切实际。

1. A serological investigation by means of an enzyme immuno assay for herpes B virus (Cercopithecine herpesvirus 1) was performed on 962 sera of healthy nonhuman primates reared in laboratory animal facilities which belong to Japanese Association of Laboratory Animal Facility of National University. An antibody prevalence of 40% (384/961) was demonstrated. The antibody titer was shown to be higher among macaques (60% of cynomolgus monkeys, 53% of rhesus monkeys, and 34% of Japanese monkeys) than among non-macaque species (21%). These data were also alarmed the use of Japanese monkeys in a research field of life science.2. We examined the use of alternative or substitutive antigen for immuno assay method instead of dangerous B virus ; 1) human herpes virus 1 (HSV1), 2) herpesvirus papio 2 (HVP2) antigen. The correlationship between HSV1 (gP and the gD antigen) and B virus ELISA was 96%. On the other hand, the correlationship between HVP2 antigen and B virus ELISA was of 98% using 350 monkey serum samples from USA and Japan. These were suggestive data that alternative method may use for the antibody assay, especially HVP2 antigen.3. It was possible to detect B virus genome in 3 samples in the antibody positive 21 sample (trigeminal ganglion) of Japanese monkeys which are originated from Japan, after trying the RT-nested PCR method.4. It accomplished in using two kinds of glycoprotein antigens of BV as the recombinant antigen. However, their products showed cytotoxic effect to mammal cells and therefore judged impractical.