Authors:Sun Xiuli, Gu Yixuan, Zhu Lei,Tian Lei, Wang Yinghui,Jie Ying
Corresponding author:Jie Ying, Email:jie_yingcn@aliyun.com
Published:2026-08-10
DOI:10.3760/cma.j.cn115989-20250916-00300
ABSTRACT
Objective To evaluate the accuracy and repeatability of fluorescence PCR in detecting TGFBI gene mutation in patients with corneal dystrophy.
Methods ARMS-PCR combined with TaqMan probe method was used for genotyping the R124H, R124C, R555W, and R555Q loci (covering heterozygous and homozygous mutation) in the TGFBI gene in peripheral blood samples from 240 patients with corneal dystrophy who were treated at Beijing Tongren Hospital, Capital Medical University from January 2021 to August 2025, and the results were compared with Sanger sequencing results. The intra-batch and inter-batch test results of fluorescence PCR were analyzed to evaluate their repeatability. The association between genotypes and clinical phenotypes was analyzed. This study adhered to the principles of the Declaration of Helsinki. The study protocol was approved by the Ethics Committee of Beijing Tongren Hospital, Capital Medical University (No. TREC2023012), and written informed consent was obtained from all patients.
Results The positive rate of pathogenic gene mutations detected by the human TGFBI gene mutation detection kit (fluorescence PCR method) and Sanger sequencing in 240 samples was 83.33%(200/240). The positive rates of R124H heterozygous mutation, R124H homozygous mutation, R124L heterozygous mutation, R124C heterozygous mutation, R555W heterozygous mutation and R555Q heterozygous mutation were 55.00%(132/240), 0.83%(2/240), 1.67%(4/240), 11.67%(28/240), 8.33%(20/240), 5.83%(14/240), respectively. The total positive rate was 83.33%(200/240), and the wild-type detection rate was 16.67%(40/240). The total coincidence rate of the two methods was 100.00%. The repeated results of intra-batch and inter-batch for TGFBI gene mutation detected by fluorescence PCR were completely consistent. Analysis of genotypes and clinical phenotypes in 200 patients with positive gene mutations showed that the differences in the distribution of clinical phenotypes among patients with different genotypes was statistically significant (χ2=498.474, P<0.001). Cramer V=0.706.
Conclusions Fluorescence PCR is highly accurate and reproducible in detecting TGFBI gene hot mutation, which is suitable for detecting gene mutation in patients with corneal dystrophy in clinical practice.
KEYWORDS：
Corneal dystrophy;TGFBI gene ;Fluorescence PCR;Sanger sequencing;Accuracy;Repeatability
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Authors Info & Affiliations
Sun Xiuli
Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Key Laboratory of Ophthalmology and Visual Science, Beijing 100730, China
Gu Yixuan
Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Key Laboratory of Ophthalmology and Visual Science, Beijing 100730, China
Zhu Lei
Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Key Laboratory of Ophthalmology and Visual Science, Beijing 100730, China
Tian Lei
Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Key Laboratory of Ophthalmology and Visual Science, Beijing 100730, China
Wang Yinghui
Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Key Laboratory of Ophthalmology and Visual Science, Beijing 100730, China
Jie Ying
Beijing Institute of Ophthalmology, Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Key Laboratory of Ophthalmology and Visual Science, Beijing 100730, China
Figures & Tables
Figure 1 Amplification curve of reference substance wt: wild type; GAPDH: glyceraldehyde-3-phos phate dehydrogenase
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