Study on the method of oocyst elution and nested PCR detection of Cryptosporidium contaminated with lettuce
LI Jun-qiang1,2, ZHANG Kai-hui1, CUI Zhao-hui1, WANG Rong-jun1, JIAN Fu-chun1, ZHANG Su-mei1, NING Chang-shen1, ZHANG Long-xian1
1. College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450046, China; 2. Academy of Chinese Medical Sciences, Henan University of Chinese Medicine, Zhengzhou 450046, China
Abstract:Cryptosporidium is an important zoonotic intestinal protozoa, which has leading caused many food-borne and water-borne cryptosporidiosis outbreaks worldwide. In order toompare the elution and detection methods for Cryptosporidium oocysts contaminated on lettuce, the elution times, oscillation patterns, oscillation frequency, oscillation time, and elution solution of Cryptosporidium oocysts by artificial ‘seeding’ on vegetable surface were compared, and three commercialized DNA extraction kits were compare used to extract oocyst DNAs. The results showed that the overall recovery rate was 71.40% when the oscillation frequency was 100 times/min, the oscillation time was 1 h, and the elution solution was 1% Alconox○R, which had the highest overall oocysts recovery rate. The higher DNAs in quality and quantity of the Cryptosporidium oocyst were obtained between soil DNA extraction kit and stool DNA extraction kit compared with tissue DNA extraction kit. And, the soil DNA kit is better if considering the time cost, and stool DNA kit is better if considering the economic cost. In conclusion, in this study we develop a nested PCR detection method for Cryptosporidium oocysts on lettuce surface, which provided a new detection method for food safety of vegetables.
李俊强,张凯慧,崔朝辉,王荣军,菅复春,张素梅,宁长申,张龙现. 生菜污染隐孢子虫卵囊洗脱与巢式PCR检测方法的研究[J]. 中国人兽共患病学报, 2020, 36(3): 211-217.
LI Jun-qiang, ZHANG Kai-hui, CUI Zhao-hui, WANG Rong-jun, JIAN Fu-chun, ZHANG Su-mei, NING Chang-shen, ZHANG Long-xian. Study on the method of oocyst elution and nested PCR detection of Cryptosporidium contaminated with lettuce. Chinese Journal of Zoonoses, 2020, 36(3): 211-217.
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