Vol.:(0123456789) 1 3
Chemical Papers
https://doi.org/10.1007/s11696-019-00948-x
ORIGINAL PAPER
Point mutation in the TGFBI gene: surface‑enhanced infrared
absorption spectroscopy (SEIRAS) as an analytical method
D. Rosas‑Vara
1
· J. R. Molina‑Contreras
1
· F. Villalobos‑Piña
1
· J. C. Zenteno
3
· B. Buentello‑Volante
3
·
O. F. Chacon‑Camacho
3
· R. Ayala‑Ramírez
3
· C. Frausto‑Reyes
4
· R. Hernández‑Martínez
2
· M. A. Ríos‑Corripio
2
Received: 15 July 2019 / Accepted: 25 September 2019
© Institute of Chemistry, Slovak Academy of Sciences 2019
Abstract
A novel method to detect DNA mutations based on gold nanoparticles is described. This bioconjugate was prepared by
conjugation of the TGFBI gene on the surface of gold nanoparticles. The surface plasmon resonance band observed in the
ultraviolet–visible spectrum of the gold nanoparticles showed a shift to a lower energy after the surface was covered with
the TGFBI gene. Surface-enhanced infrared spectroscopy (SEIRAS) showed the absorption bands of the bioconjugate due
to their proximity to the nanoparticles. The SEIRAS efect was able to detect small and specifc changes in the sequence
of the TGFBI gene using interactions of the bioconjugates. The study was performed on patients clinically diagnosed with
lattice corneal dystrophy (LCD). To our knowledge, this is the frst report of FTIR spectroscopy employed to analyse this
gene. We used genotyping results previously obtained by next-generation DNA sequencing technology to ensure that the
analysed samples had the mutation. Our results show highly reproducible signals, which could be used for analytical studies
of mutations in the genome.
Keywords FTIR · DNA · PCR · Lattice corneal dystrophy (LCD) · Oculopharyngeal muscular dystrophy (OPMD)
Introduction
Knowledge of the existence of DNA mutations may aid
in the timely treatment of diseases. Human transforming
growth factor β-induced (TGFBI also known as BIGH3), is
a gene responsible for various corneal dystrophies. TGFBI
produces a protein called TGFBI, which is involved in cell
adhesion and serves as a recognition sequence for integ-
rins. An alteration in cell surface interactions could be
the underlying cause for the progressive accumulation of
extracellular deposits in diferent layers of the cornea with
the resulting changes of refractive index and transparency.
To this date, 69 diferent pathogenic or likely pathogenic
variants in TGFBI have been identifed in a heterozygous or
homozygous state in various corneal dystrophies (Kheir et al.
2019). For this reason, efcient and cost-efective mutation
detection technologies are of paramount importance for both
research and the progress of molecular genetics. A variety
of methods for identifying mutations have been published
(Chin et al. 2013; Harada and Korf 2013; Southern 1996).
However, many of these methods and techniques are time
consuming, and some of them cannot identify structural and
compositional changes in the samples simultaneously. This
shows the need to implement new methodologies that would
enable a reliable screening diagnosis within a short time. It
is at this point where spectroscopy appears as a modern and
attractive technique in the study of DNA and the detection
of mutations. Infrared (IR) spectroscopy provides a unique
vibrational phenotypic fngerprint that is complementary to
genomic approaches to distinguish the allelic forms of each
gene. Attenuated total refection Fourier transform infra-
red spectroscopy (ATR-FTIR) is one of the most powerful
* M. A. Ríos-Corripio
anto200784@yahoo.com.mx
1
Department of Basic Sciences, Instituto Tecnológico de
Aguascalientes, Aguascalientes, Mexico
2
CONACYT-Colegio de Postgraduados-Campus Córdoba,
Carretera federal Córdoba-Veracruz Km. 348, Amatlán de los
Reyes, C.P. 94946 Veracruz, Mexico
3
Department of Genetics, Research Unit, Instituto de
Oftalmología “Conde de Valenciana” and Department
of Biochemistry, Faculty of Medicine, UNAM, México, DF,
Mexico
4
A.C., Aguascalientes Unit, Center for Research in Optics,
Aguascalientes, Mexico