RT Journal Article
T1 Novel molecular diagnostic system of limbal stem cell deficiency based on MUC5AC transcript detection in corneal epithelium by PCR-reverse dot blot
A1 García, Iker
A1 Etxebarria, Jaime
A1 Merayo Lloves, Jesús
A1 Torras, Josep
A1 Boto de los Bueis, Ana
A1 Díaz Valle, David
A1 Méndez Fernández, Rosalía
A1 Acera, Arantxa
A1 Suárez, Tatiana
AB Purpose. To evaluate the sensitivity and specificity of a PCR-strip system based on reverse dot blot for detection of MUC5AC mRNA in corneal epithelium samples from patients with limbal stem cell deficiency (LSCD), and to determine the correlation with clinical diagnosis. Methods. We obtained 87 corneal impression cytology (IC) samples from 55 subjects (37 patients clinically diagnosed with LSCD and 18 control subjects). Total RNA was extracted from each IC sample and retrotranscribed to cDNA. MUC5AC mRNA transcript was amplified by a customized RT-PCR assay and detected in PCR strips based on reverse dot blot hybridization and in agarose gels. Conjunctival IC samples were used as positive controls. Results. Forty-four of 45 corneal IC samples obtained from patients clinically diagnosed with LSCD were positive for MUC5AC, whereas 34 of 42 corneal ICs from healthy subjects were negative for MUC5AC. Four healthy corneas were found MUC5AC positive, and four rendered inconclusive results. A correlation of 91.4% (P < 0.001) between molecular testing and clinical diagnosis was found. The overall sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the PCR-strip system were 98%, 89%, 92%, and 97%, respectively. Conclusions. Corneal epithelium MUC5AC transcript detection by reverse dot blot PCR-strip is a highly sensitive technique for LSCD diagnosis. The test results strongly correlate with clinical diagnosis of characterized LSCD cases. The PCR-strip system may be used for early detection, and for the detection of mild cases of LSCD, and constitutes an objective clinical tool for the monitoring of treatments and surgical decisions
PB The Association for Research in Vision and Ophthalmology
SN 1552-5783
YR 2013
FD 2013-08-10
LK https://hdl.handle.net/20.500.14352/112287
UL https://hdl.handle.net/20.500.14352/112287
LA eng
NO : García I, Etxebarria J, MerayoLloves J, et al. Novel molecular diagnostic system of limbal stem cell deficiency based on MUC5AC transcript detection in corneal epithelium by PCR-reverse dot blot. Invest Ophthalmol Vis Sci. 2013;54:5643–5652. DOI:10.1167/iovs.13-11933
NO Centre for the Development of Industrial Technology
DS Docta Complutense
RD 7 abr 2025