Real-Time Reverse Transcription Quantitative PCR (RT-qPCR) Methodological Standards and Reporting Practices

Reporting of core parameters remained low or declined. Between 2019 and 2024, RNA integrity reporting fell (22% to 11%), reference gene validation was rare (13% to 5%), and PCR efficiency reporting collapsed (13% to 1%). MIQE-citing papers in 2024 showed better adherence (31% RNA integrity, 47% reference gene validation, and 40% PCR efficiency) but still omitted essential details. Asia now dominates RT-qPCR output by volume, while Europe contributes most MIQE-citing studies. Targeted cohorts reported more methodological information, yet many still failed to meet basic standards. These findings confirm that incomplete experimental design and reporting continue to undermine reproducibility and robustness of RT-qPCR assays.