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在检测牛淋巴结时,有内对照和无内对照的双靶点实时荧光定量PCR检测方法的比较数据。

Comparison data of a two-target real-time PCR assay with and without an internal control in detecting from cattle lymph nodes.

作者信息

Bai Jianfa, Trinetta Valentina, Shi Xiaorong, Noll Lance W, Magossi Gabriela, Zheng Wanglong, Porter Elizabeth P, Cernicchiaro Natalia, Renter David G, Nagaraja Tiruvoor G

机构信息

Kansas State Veterinary Diagnostic Laboratory, College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506, Unites States.

Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS 66506, United States.

出版信息

Data Brief. 2018 Apr 22;18:1819-1824. doi: 10.1016/j.dib.2018.04.051. eCollection 2018 Jun.

Abstract

A real-time PCR (qPCR) assay targeting on A and C genes was developed and validated for the detection and quantification of strains (Bai et al., 2018) [1]. A host gene, normally an endogenous housekeeping gene (Beer-Davidson et al., 2018; Poon et al., 2004) [2,3], or an irrelevant exogenous gene (Cheng et al., 2015; Sedlak et al., 2014) [4,5] has been widely used as an internal control to monitor nucleic acid extraction efficiencies and potential PCR inhibitions in PCR-based detection assays. An endogenous internal control designed based on the 18S rRNA gene was used in the above-mentioned qPCR assay. This 18S rRNA internal control amplifies the target gene in multiple species including bovine, swine, ovine, caprine and cervine. Data was generated by the duplex qPCR assay on 138 enriched cattle lymph node samples without the internal control, and compared with data on the same samples tested by the triplex qPCR assay that has the 18S rRNA gene as internal control. Threshold cycle (Ct) data for the duplex and the triplex qPCR on the 138 samples were similar, and are presented in this brief report.

摘要

开发了一种针对A和C基因的实时荧光定量PCR(qPCR)检测方法,并进行了验证,用于菌株的检测和定量(白等人,2018年)[1]。宿主基因,通常是内源性管家基因(比尔 - 戴维森等人,2018年;潘等人,2004年)[2,3],或无关的外源基因(程等人,2015年;塞德拉克等人,2014年)[4,5]已被广泛用作内部对照,以监测基于PCR的检测方法中的核酸提取效率和潜在的PCR抑制作用。上述qPCR检测方法中使用了基于18S rRNA基因设计的内源性内部对照。这种18S rRNA内部对照可在包括牛、猪、羊、山羊和鹿在内的多个物种中扩增目标基因。通过对138个未添加内部对照的富集牛淋巴结样本进行双重qPCR检测生成数据,并与通过以18S rRNA基因为内部对照的三重qPCR检测相同样本的数据进行比较。138个样本的双重和三重qPCR的阈值循环(Ct)数据相似,本简要报告中呈现了这些数据。

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