Comparison of Automated and Manual DNA Isolation Methods of Liquid-Based Cytology Samples
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 procedur...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Mary Ann Liebert, Inc.
2019
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| Subjects: | |
| Online Access: | http://eprints.uanl.mx/27189/7/27189.pdf.crdownload |
| _version_ | 1824420679144964096 |
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| author | Domínguez Vigil, Irma Guadalupe Barajas Olmos, Víctor H. Gallardo Alvarado, Lenny Pérez Maya, Antonio Alí Garza Rodríguez, María L. Magallanes Garza, Gerardo I. Cardona Huerta, Servando Méndez Lozano, Daniel H. Vidal Gutiérrez, Oscar Cantú De León, David F. Barrera Saldaña, Hugo Alberto |
| author_facet | Domínguez Vigil, Irma Guadalupe Barajas Olmos, Víctor H. Gallardo Alvarado, Lenny Pérez Maya, Antonio Alí Garza Rodríguez, María L. Magallanes Garza, Gerardo I. Cardona Huerta, Servando Méndez Lozano, Daniel H. Vidal Gutiérrez, Oscar Cantú De León, David F. Barrera Saldaña, Hugo Alberto |
| author_sort | Domínguez Vigil, Irma Guadalupe |
| collection | Repositorio Institucional |
| description | 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. |
| format | Article |
| id | eprints-27189 |
| institution | UANL |
| language | English |
| publishDate | 2019 |
| publisher | Mary Ann Liebert, Inc. |
| record_format | eprints |
| spelling | eprints-271892024-12-11T16:47:51Z http://eprints.uanl.mx/27189/ Comparison of Automated and Manual DNA Isolation Methods of Liquid-Based Cytology Samples Domínguez Vigil, Irma Guadalupe Barajas Olmos, Víctor H. Gallardo Alvarado, Lenny Pérez Maya, Antonio Alí Garza Rodríguez, María L. Magallanes Garza, Gerardo I. Cardona Huerta, Servando Méndez Lozano, Daniel H. Vidal Gutiérrez, Oscar Cantú De León, David F. Barrera Saldaña, Hugo Alberto RG Ginecología y Obstetricia 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. Mary Ann Liebert, Inc. 2019 Article PeerReviewed text en cc_by_nc_nd http://eprints.uanl.mx/27189/7/27189.pdf.crdownload http://eprints.uanl.mx Domínguez Vigil, Irma Guadalupe y Barajas Olmos, Víctor H. y Gallardo Alvarado, Lenny y Pérez Maya, Antonio Alí 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 Alberto (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 http://doi.org/10.1089/bio.2018.0148 |
| spellingShingle | RG Ginecología y Obstetricia Domínguez Vigil, Irma Guadalupe Barajas Olmos, Víctor H. Gallardo Alvarado, Lenny Pérez Maya, Antonio Alí Garza Rodríguez, María L. Magallanes Garza, Gerardo I. Cardona Huerta, Servando Méndez Lozano, Daniel H. Vidal Gutiérrez, Oscar Cantú De León, David F. Barrera Saldaña, Hugo Alberto Comparison of Automated and Manual DNA Isolation Methods of Liquid-Based Cytology Samples |
| thumbnail | https://rediab.uanl.mx/themes/sandal5/images/online.png |
| title | Comparison of Automated and Manual DNA Isolation Methods of Liquid-Based Cytology Samples |
| title_full | Comparison of Automated and Manual DNA Isolation Methods of Liquid-Based Cytology Samples |
| title_fullStr | Comparison of Automated and Manual DNA Isolation Methods of Liquid-Based Cytology Samples |
| title_full_unstemmed | Comparison of Automated and Manual DNA Isolation Methods of Liquid-Based Cytology Samples |
| title_short | Comparison of Automated and Manual DNA Isolation Methods of Liquid-Based Cytology Samples |
| title_sort | comparison of automated and manual dna isolation methods of liquid based cytology samples |
| topic | RG Ginecología y Obstetricia |
| url | http://eprints.uanl.mx/27189/7/27189.pdf.crdownload |
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