Application of Duplex Droplet Digital PCR Detection of miR-888 and miR-891a in Semen Identification / 法医学杂志
Journal of Forensic Medicine
; (6): 719-725, 2022.
Article
ي En
| WPRIM
| ID: wpr-984163
المكتبة المسؤولة:
WPRO
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.
Key words
النص الكامل:
1
الفهرس:
WPRIM
الموضوع الرئيسي:
Saliva
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Semen
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Body Fluids
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MicroRNAs
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Real-Time Polymerase Chain Reaction
المحددات:
Female
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Humans
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Male
اللغة:
En
مجلة:
Journal of Forensic Medicine
السنة:
2022
نوع:
Article