基孔肯雅病毒感染期间合适管家基因的鉴定

Identification of suitable house-keeping genes during chikungunya virus infection.

作者信息

Agrawal Nishtha, Khanna Madhu, Dhawan Gagan

机构信息

Department of Virology (a Unit of Dept. of Microbiology), Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, 110007, India; Department of Biomedical Science, Acharya Narendra Dev College, University of Delhi, Kalkaji, New Delhi, 110019, India.

Department of Virology (a Unit of Dept. of Microbiology), Vallabhbhai Patel Chest Institute, University of Delhi, Delhi, 110007, India.

出版信息

Indian J Med Microbiol. 2023 Mar-Apr;42:49-52. doi: 10.1016/j.ijmmb.2023.01.007. Epub 2023 Feb 3.

DOI:10.1016/j.ijmmb.2023.01.007

PMID:36967216

Abstract

PURPOSE

Quantitative PCR (qPCR) is a reliable and robust technique for gene expression analysis, but its efficacy is dependent on the normalization of qPCR data with the stably expressed reference gene. Selection of a suitable reference gene is mandatory for accurate gene expression analysis, till data the most appropriate reference gene during chikungunya virus infection has not been elucidated.

METHOD

In this study the expression of reference genes(GAPDH, GUSB, HPRT, Beta-actin, 18S rRNA) was analysed during chikungunya virus infection by quantitative PCR. The stability of the house-keeping genes was evaluated with three bioinformatics softwares: BestKeeper, NormFinder and GeNorm.

RESULT

The significant variation in the expression of house-keeping genes (GusB, Beta-actin, HPRT) was observed during chikungunya virus infection; whereas GAPDH and 18S rRNA was most stable. The stability of reference genes analysed by the bioinformatics software further corroborate the results of qPCR.

CONCLUSION

This is first study that identifies and validates the most suitable reference gene for normalization of qPCR data during chikungunya based gene expression analysis. This could serve as a reference study for the researchers working on different aspects of chikungunya virus infections.

摘要

目的

定量聚合酶链反应（qPCR）是一种用于基因表达分析的可靠且强大的技术，但其有效性取决于使用稳定表达的参考基因对qPCR数据进行标准化。选择合适的参考基因对于准确的基因表达分析至关重要，然而，在基孔肯雅病毒感染期间最合适的参考基因尚未明确。

方法

在本研究中，通过定量PCR分析了基孔肯雅病毒感染期间参考基因（甘油醛-3-磷酸脱氢酶（GAPDH）、β-葡萄糖醛酸酶（GUSB）、次黄嘌呤磷酸核糖转移酶（HPRT）、β-肌动蛋白、18S核糖体RNA）的表达。使用三种生物信息学软件（BestKeeper、NormFinder和GeNorm）评估管家基因的稳定性。