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Gaceta médica de México
versión On-line ISSN 2696-1288versión impresa ISSN 0016-3813
Resumen
DIMAS-GONZALEZ, Jisela; TREJO-GOMORA, Jorge E.; BONILLA-CISNEROS, Carlos y LAGUNAS-MARTINEZ, Alfredo. Colony-forming units optimization and human papillomavirus detection in umbilical cord blood. Gac. Méd. Méx [online]. 2021, vol.157, n.1, pp.30-36. Epub 18-Jun-2021. ISSN 2696-1288. https://doi.org/10.24875/gmm.19005696.
Introduction:
Analysis of several markers is required for adequate quality control in umbilical cord blood units (UCBU) when are used for therapeutic purposes.
Objective:
To optimize colony-forming units (CFU) from clonogenic cultures and to detect the human papillomavirus (HPV) genome in UCBU.
Methods:
One hundred and forty-one umbilical cord blood (UCB), segment or CFU samples from UCBU clonogenic cultures were included. DNA extraction, quantification and endogenous GAPDH gene PCR amplification were carried out. Subsequently, HPV L1 gene was detected using the MY09/MY11 and GP5/GP6+ oligonucleotides. PCR products were analyzed with electrophoresis in agarose gel. CFU-extracted purified DNA was analyzed by electrophoresis in agarose gel, as well as some DNAs, using the SSP technique.
Results:
CFU-extracted DNA concentration was higher in comparison with that of UCB (p = 0.0041) and that of the segment (p < 0.0001), as well as that of UCB in comparison with that of the segment (p < 0.0001). All samples were positive for GAPDH amplification and negative for MY09/MY11 and GP5/GP6+.
Conclusions:
Cryopreserved UCBUs were HPV-negative. Obtaining CFU DNA from clonogenic cultures with high concentrations and purity is feasible.
Palabras llave : Human papillomavirus; Clonogenic cultures; Colony-forming units; Umbilical cord blood units; DNA.