Identification of transcriptional biomarkers by RNA-sequencing for improved detection of β2-agonists abuse in goat skeletal muscle

PLoS One. 2017 Jul 26;12(7):e0181695. doi: 10.1371/journal.pone.0181695. eCollection 2017.

Abstract

In this paper, high-throughput RNA-sequencing (RNA-seq) was used to search for transcriptional biomarkers for β2-agonists. In combination with drug mechanisms, a smaller group of genes with higher detection accuracy was screened out. Unknown samples were first predicted by this group of genes, and liquid chromatograph tandem mass spectrometer (LC-MS/MS) was applied to positive samples to validate the biomarkers. The results of principal component analysis (PCA), hierarchical cluster analysis (HCA) and discriminant analysis (DA) indicated that the eight genes screened by high-throughput RNA-seq were able to distinguish samples in the experimental group and control group. Compared with the nine genes selected from an earlier literature, 17 genes including these nine genes were proven to have a more satisfactory effect, which validated the accuracy of gene selection by RNA-seq. Then, six key genes were selected from the 17 genes according to the variable importance in projection (VIP) value of greater than 1. The test results using the six genes and 17 genes were similar, revealing that the six genes were critical genes. By using the six genes, three positive samples possibly treated with drugs were screened out from 25 unknown samples through DA and partial least squares discriminant analysis (PLS-DA). Then, the three samples were verified by a standard method, and mapenterol was detected in a sample. Therefore, the six genes can be used as biomarkers to detect β2-agonists. Compared with the previous study, accurate detection of β2-agonists abuse using six key genes is an improvement method, which show great significance in the monitoring of β2-agonists abuse in animal husbandry.

MeSH terms

  • Adrenergic beta-Agonists / metabolism
  • Adrenergic beta-Agonists / pharmacology*
  • Animals
  • Biomarkers / analysis*
  • Chromatography, Liquid
  • Cluster Analysis
  • Discriminant Analysis
  • Gene Expression / drug effects*
  • Goats
  • Least-Squares Analysis
  • Male
  • Muscle, Skeletal / drug effects*
  • Muscle, Skeletal / metabolism
  • Principal Component Analysis
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Analysis, RNA / methods*
  • Substance Abuse Detection / methods
  • Substance Abuse Detection / veterinary*
  • Tandem Mass Spectrometry

Substances

  • Adrenergic beta-Agonists
  • Biomarkers

Grants and funding

This work is supported by Special Fund for Agro-scientific Research in the Public Interest 201203023.