The Vaginal Microbiota Analysis by Polymerase Chain Reaction-high-resolution Melting
Jinling Li, Menglu Chen, Mengfen Mao, Tuerxun Adilai, Haojie Qin, Wei Feng, Yaonan Mo, Xiandun ZhaiBackground:
The vaginal microbiota is individually characteristic and forensically promising, yet current detection methods are often too time-consuming or costly for rapid preliminary screening.
Aims:
The vaginal microbiota from healthy and reproductive-age women was analyzed based on polymerase chain reaction-high resolution melting curve (PCR-HRM) to provide basic data for forensic practice.
Methods:
100 vaginal secretions from healthy and reproductive-age women who met the inclusion criteria were collected. As the target fragment, the
Results:
The results of the positive control and the predicted profiles were completely consistent. Consistent profiles of multiple samples could be obtained from repeated and parallel experiments. Effective amplification curves could still be obtained when the concentration of samples is 0.15 ng/µL, but no amplification curve was found from human DNA and the negative control. According to the different shapes of melting curves and Tm value, 100 samples in this study were finally classified into 10 types. The bacteria showing dominant peaks in the profiles were
Conclusion:
The method for detecting female vaginal microbiota based on PCR-HRM has the advantage of high sensitivity, good species specificity, good repeatability, and high stability. In some special cases, a large number of samples of vaginal secretions can be preliminarily screened and classified with the method in a short time, which can be used as a supplementary means to obtain evidence of crimes of individual identification and source of body fluid through multiple means and dimensions.