Kassouha Morshed, Soukkarieh Chadi, Alkhaled Abdulkarim
Department of Microbiology, Faculty of Veterinary Medicine, Hama University, Hama, Syria.
Department of Animal Biology, Faculty of Sciences, Damascus University, Damascus, Syria.
Vet Parasitol. 2016 Jul 30;225:86-90. doi: 10.1016/j.vetpar.2016.06.009. Epub 2016 Jun 6.
In this study, PCR-RFLP was used for the first time in Syria for genotyping Cryptosporidium species of man, calves and chickens. The total of 391 fecal samples included 213 from children with diarrhea (<5years), 67 from pre-weaned calves with diarrhea and 111 from broiler chicken farms. All samples were collected and examined with acid fast stain to detect the positive samples. Subsequently a nested-PCR test was performed on 35 positive samples (17 from calves, 11 from chicken, and 7 from children) targeting SSU rRNA gene, and was followed by RFLP analysis using three restriction enzymes SspI, VspI and MboII. Results showed that C. parvum was the only identified species in children and calves, on the other hand C. baileyi was identified in broilers in addition to another species with unknown RFLP profile in comparison to those which have been described in chicken. Further studies using more genes are needed to sequence and detect subtypes of this parasite.
在本研究中，聚合酶链反应-限制性片段长度多态性分析（PCR-RFLP）首次在叙利亚用于对人类、小牛和鸡的隐孢子虫物种进行基因分型。总共391份粪便样本包括213份来自腹泻儿童（<5岁）、67份来自腹泻的断奶前小牛以及111份来自肉鸡养殖场。所有样本均收集并用抗酸染色法检查以检测阳性样本。随后，对35份阳性样本（17份来自小牛、11份来自鸡、7份来自儿童）进行巢式PCR检测，靶向小亚基核糖体RNA（SSU rRNA）基因，接着使用三种限制性内切酶SspI、VspI和MboII进行RFLP分析。结果表明，微小隐孢子虫是在儿童和小牛中唯一鉴定出的物种，另一方面，除了与已在鸡中描述的具有不同RFLP图谱的另一个物种外，在肉鸡中鉴定出了贝氏隐孢子虫。需要使用更多基因进行进一步研究以对该寄生虫的亚型进行测序和检测。
