Comparison of Automated and Manual DNA Isolation Methods of Liquid-Based Cytology Samples

Domínguez Vigil, Irma G. y Barajas Olmos, Víctor H. y Gallardo Alvarado, Lenny y Pérez Maya, Antonio A. y Garza Rodríguez, María L. y Magallanes Garza, Gerardo I. y Cardona Huerta, Servando y Méndez Lozano, Daniel H. y Vidal Gutiérrez, Oscar y Cantú De León, David F. y Barrera Saldaña, Hugo A. (2019) Comparison of Automated and Manual DNA Isolation Methods of Liquid-Based Cytology Samples. Biopreservation and Biobanking, 17 (6). pp. 591-597. ISSN 1947-5535

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Resumen

Liquid-based cytology (LBC) has been used as a diagnostic tool for cervical cancer for years and is now being adopted for other gynecological cancers. LBC represents an important challenge to ensure that the process yields representative biospecimens for quality control (QC) of diagnostic procedures. In this study, we compare QC parameters (integrity, yield and purity, and polymerase chain reaction [PCR] amplification) of DNA isolated from LBC (N= 296) using two different nucleic acid isolation methods, manual (n = 233) or automated (n = 63). We also evaluated two different types of cytological brushes for sampling from the cervix. Our results suggest that manual isolation (yield 22.81 – 1.92 mg) resulted in increased DNA recovery when compared with automated isolation (yield 9.96 – 1.11 mg) from LBC samples, with a p-value of <0.0003. We estimated that 98% (53/54) of the samples preserved the integrity of DNA and were suitable for standard molecular biology analyses. The b-globin gene was amplified in 100% (296/296) of the DNA samples by endpoint PCR. We found no significant difference between the performance of the cytological brushes ( p value of <0.6711) in a general overview. However, when looking at the results from using each brush individually, the manual isolation method was statistically superior to the automated method. Our work illustrates the impact of good QC of preanalytic conditions, which will be important for the application of LBC for developing early detection methods for gynecological cancers.

Tipo de elemento:

Article

Palabras claves no controlados:

Cáncer de cuello uterino; Citología líquida; Ácidos nucleicos; Control de calidad; Biobanco; Biomarcador.

Materias:

R Medicina > RG Ginecología y Obstetricia

Divisiones:

Medicina

Usuario depositante:

Dr Irma G Dominguez Vigil

Creadores:

Creador	Email	ORCID
Domínguez Vigil, Irma G.	irma52vigil@gmail.com	orcid.org/0000-0003-4687-6318
Barajas Olmos, Víctor H.	NO ESPECIFICADO	NO ESPECIFICADO
Gallardo Alvarado, Lenny	NO ESPECIFICADO	NO ESPECIFICADO
Pérez Maya, Antonio A.	NO ESPECIFICADO	NO ESPECIFICADO
Garza Rodríguez, María L.	NO ESPECIFICADO	NO ESPECIFICADO
Magallanes Garza, Gerardo I.	NO ESPECIFICADO	NO ESPECIFICADO
Cardona Huerta, Servando	NO ESPECIFICADO	NO ESPECIFICADO
Méndez Lozano, Daniel H.	NO ESPECIFICADO	NO ESPECIFICADO
Vidal Gutiérrez, Oscar	NO ESPECIFICADO	NO ESPECIFICADO
Cantú De León, David F.	NO ESPECIFICADO	NO ESPECIFICADO
Barrera Saldaña, Hugo A.	habarrera@gmail.com	orcid.org/0000-0003-1587-4571