Ghorbani Marghmaleki Elahe, Ahmadi Azam, Arjomandzadegan Mohammad, Akbari Majid, Karamghoshchi Aysan
Infectious Diseases Research Center (IDRC), Arak University of Medical Sciences, Arak, Iran.
Rep Biochem Mol Biol. 2020 Oct;9(3):257-263. doi: 10.29252/rbmb.9.3.257.
spp. are the main cause of human gastroenteritis. The sequencing is one of fast molecualr method to detect this fastidious. In this study, we compared the sequencing of genewith four housekeeping genestodetect spp. in patients with diarrhea and healthy people.
60 samples of DNA extracted from stool samples of 30 patients with diarrhea and 30 healthy people were used. In order to detect , we designed primers for proliferation of , and genes using Primer 3, Mega 4.0 and Blast software. Then the PCR products were sequenced using ABI system.
The sequencing showed concordance of PCR-products with deposited sequences in the Gene Bank. Among diarrhea patients, 53.3% of samples were significantly (p< 0.05) positive for and genes and 50% of samples were positive using , and genes by PCR assay. The average of sensitivity and specificity were found 53.33% and 83.33%, respectively.
Due to various copies of repeated sequences of gene, analyzing its amplicons on electrophoresis may be more difficult than the and genes. According to our results, among the 5 studied genes; the highest detection rate was related to and genes. Although, and genes,instead of gene, can be considered as appropriate genes for molecular detection of bacteria.
[具体物种名称]是人类肠胃炎的主要病因。[具体基因名称]测序是检测这种苛求菌的快速分子方法之一。在本研究中，我们比较了[具体基因名称]基因测序与四个管家基因测序，以检测腹泻患者和健康人群中的[具体物种名称]。
使用从30例腹泻患者和30名健康人的粪便样本中提取的60份DNA样本。为了检测[具体物种名称]，我们使用Primer 3、Mega 4.0和Blast软件设计了用于扩增[具体基因名称1]、[具体基因名称2]和[具体基因名称3]基因的引物。然后使用ABI系统对PCR产物进行测序。
测序结果显示PCR产物与基因库中保存的序列一致。在腹泻患者中，53.3%的样本对[具体基因名称1]和[具体基因名称2]基因呈显著阳性（p<0.05），50%的样本通过PCR检测对[具体基因名称3]、[具体基因名称4]和[具体基因名称5]基因呈阳性。敏感性和特异性的平均值分别为53.33%和83.33%。
由于[具体基因名称]基因重复序列的拷贝数不同，在电泳上分析其扩增子可能比[具体基因名称1]和[具体基因名称2]基因更困难。根据我们的结果，在研究的5个基因中，最高检测率与[具体基因名称1]和[具体基因名称2]基因相关。尽管如此，[具体基因名称3]和[具体基因名称4]基因而非[具体基因名称]基因可被视为用于[具体细菌名称]分子检测的合适基因。
