微生物农药耳霉菌致病性的PCR检测

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目的应用PCR扩增技术对微生物农药耳霉菌经口给予大鼠可能引起的感染性和致病性进行评价。方法将50只健康7~9周龄SPF级Wistar大鼠按体重随机分为两组,分别为对照组10只,实验组40只,雌雄各半。将1.0×108units/ml微生物农药耳霉菌悬浮剂按1.0 ml/只经口一次性灌胃给予大鼠,对照组不做处理;连续观察21 d。记录动物的体重、中毒症状、死亡情况,并对死亡动物进行大体解剖。于染毒后第0、3、7、14、21天分别处死实验组动物各6只,雌雄各半;对照组动物在第21天处死,收集不同时间点和不同组别的动物血液、脑、淋巴结、肝、脾、肺、肾等组织及粪便,提取其中的基因组DNA,进行PCR扩增实验,并对相应脏器进行病理学检测。结果各组实验动物在实验周期内无明显中毒症状,未见死亡,大体解剖及病理组织检查未见明显异常。染毒后各个时间点所取材各组动物血液及组织中耳霉菌的PCR扩增结果均为阴性。结论微生物农药耳霉菌经口急性染毒后对实验动物大鼠未见感染性及致病性。 OBJECTIVE: To evaluate the infectivity and pathogenicity caused by oral administration of Microbial Pestis fungi in rats by PCR amplification. Methods Fifty healthy Wistar rats aged 7-9 weeks were randomly divided into two groups according to body weight: 10 in the control group, 40 in the experimental group, and half in the male and the female. The 1.0 × 108units / ml microbial pesticides ear fungus suspension of 1.0 ml / oral only once by intragastric administration of rats, the control group without treatment; continuous observation of 21 d. Animals were recorded for weight, symptoms of poisoning and death, and gross anatomy of dead animals. Animals were sacrificed on the 21st day after sacrificing on days 0, 3, 7, 14 and 21. The animals in the control group were sacrificed on the 21st day to collect blood and brain of animals at different time points and in different groups , Lymph nodes, liver, spleen, lung, kidney and other stool and feces, extracted genomic DNA, PCR amplification experiments, and the corresponding pathological organ detection. Results The experimental animals in each group had no obvious symptoms of poisoning during the experimental period, no death was found, and no obvious abnormalities were found in gross anatomy and pathological examination. The results of PCR amplification of the ear molds in the blood and tissues of each group of animals taken at various time points after exposure were all negative. CONCLUSION No infection and pathogenicity were observed in experimental animals after acute oral administration of the fungus.
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