Application of Duplex Droplet Digital PCR Detection of miR-888 and miR-891a in Semen Identification.
Fa Yi Xue Za Zhi
; 38(6): 719-725, 2022 Dec 25.
Article
em En, Zh
| MEDLINE
| ID: mdl-36914387
ABSTRACT
OBJECTIVES:
To establish a system for simultaneous detection of miR-888 and miR-891a by droplet digital PCR (ddPCR), and to evaluate its application value in semen identification.METHODS:
The hydrolysis probes with different fluorescence modified reporter groups were designed to realize the detection of miR-888 and miR-891a by duplex ddPCR. A total of 75 samples of 5 body fluids (including peripheral blood, menstrual blood, semen, saliva and vaginal secretion) were detected. The difference analysis was conducted by Mann-Whitney U test. The semen differentiation ability of miR-888 and miR-891a was evaluated by ROC curve analysis and the optimal cut-off value was obtained.RESULTS:
There was no significant difference between the dual-plex assay and the single assay in this system. The detection sensitivity was up to 0.1 ng total RNA, and the intra- and inter-batch coefficients of variation were less than 15%. The expression levels of miR-888 and miR-891a detected by duplex ddPCR in semen were both higher than those in other body fluids. ROC curve analysis showed that the AUC of miR-888 was 0.976, the optimal cut-off value was 2.250 copies/µL, and the discrimination accuracy was 97.33%; the AUC of miR-891a was 1.000, the optimal cut-off value was 1.100 copies/µL, and the discrimination accuracy was 100%.CONCLUSIONS:
In this study, a method for detection of miR-888 and miR-891a by duplex ddPCR was successfully established. The system has good stability and repeatability and can be used for semen identification. Both miR-888 and miR-891a have high ability to identify semen, and the discrimination accuracy of miR-891a is higher.Palavras-chave
Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Líquidos Corporais
/
MicroRNAs
Tipo de estudo:
Diagnostic_studies
/
Prognostic_studies
Limite:
Female
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Humans
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Male
Idioma:
En
/
Zh
Revista:
Fa Yi Xue Za Zhi
Assunto da revista:
JURISPRUDENCIA
Ano de publicação:
2022
Tipo de documento:
Article
País de afiliação:
China