Identification of Suitable Reference Genes for Investigating Gene Expression in Anterior Cruciate Ligament Injury by Using Reverse Transcription-Quantitative PCR

• DOI: 10.1371/journal.pone.0133323
• Publication Date: 2015-07-20T13:51:37Z
• Authors: Mariana Ferreira ...; Diego Costa Astur; Pedro Debieux; Gustavo Gonçalves...; Carlos Eduardo Si...; Leonor Casilla Lo...; Carlos Vicente An...; Marília Cardoso S...; Alberto de Castro...; Benno Ejnisman; Moises Cohen
• Keywords: Reference Genes; ACL injury

The anterior cruciate ligament (ACL) is one of the most frequently injured structures during high-impact sporting activities. Gene expression analysis may be a useful tool for understanding ACL tears and healing failure. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) has emerged as an effective method such studies. However, this technique requires use suitable reference genes data normalization. Here, we evaluated suitability six (18S, ACTB, B2M, GAPDH, HPRT1, TBP) by using samples 39 individuals with (20 isolated 19 tear combined meniscal injury) 13 controls. stability candidate was determined NormFinder, geNorm, BestKeeper DataAssist, RefFinder software packages comparative ΔCt method. ACTB best single gene ACTB+TBP pair. GenEx showed that accumulated standard deviation reduced when larger number used not suitable. To identify optimal combination genes, FN1 PLOD1. We observed at least 3 should used. ACTB+HPRT1+18S trio analyses involving Conversely, ACTB+TBP+18S (1) controls, (2) patients Therefore, if study aims to compare non-injured ACL, from three independent groups ACTB+TBP+18S+HPRT1 In conclusion, or more without tears.