Human papilloma virus (HPV) and cervical cancer and cervical epithelioid variant (CIN) occurs in the presence of a close relationship, and according to the induced tumor difference can be divided into two kinds of low-risk and high-risk, respectively, benign tumor and malignant tumor [1] caused by certain probability. Based on this study, this paper discusses the clinical significance of multiplex real-time PCR technique for detection of high risk human papilloma virus infection. Now the report is as follows:
1, data and methods
1.1, the general data of Jiaozuo Maternal and Child Health-Care Hospital in July 2012 - December 2013 period of suspected cervical lesions and infection of HPV cases of 120 cases, the age of 28 to 71 years old, average (41.2 + 6.7) years old. Check before the start of the 3D within the vaginal drugs and vaginal flushing; check the time to avoid the menstrual period, while the 24h within the prohibition of sexual behavior.
1. 2, methods HC2 assay: the detection system is provided by the United States Di-gene company, detection project involving 13 types of high-risk HPV DNA, positive evaluation criteria [2] are as follows: sample to be measured relative light units (RLU) with the standard positive control RLU ratio exceeds the standard reference value, if the sample RLU/ standard positive control RLU > 1. 0 can be determined as positive. Multiple real-time PCR detection: the instrument used in the production of the German ABI7300 fluorescence quantitative gene amplification instrument, the test reagents are used to provide all the Shanghai blue based biological Co., ltd.. Of 13 types of high-risk HPV DNA testing, and extract the HC2 test samples, the pH was adjusted to 7. 4, take HC2 tested samples of 2 ml PCR reaction liquid evenly mixed, then by fluorescence quantitative PCR instrument were amplified and positive evaluation criteria [3] are as follows: sample cycle threshold below 37 can be judged to be positive.
1.3. PCR and HC2 were used to detect the high risk type HPV and subtype infection.
1. 4. Statistical analysis using SPSS16. 0 software and measurement data (x ~ + s) said measurement data t test was used to compare. Count data expressed as a percentage and application of 2 test, P < 0.05, the difference was statistically significant.
2, knot fruit
2. 1, high-risk HPV detection results in the detection of high-risk HPV, this study used two methods for detecting efficiency approximation, there was no statistically significant differences (P > 0. 05); compared with the wet wart and cervicitis, multiplex real-time PCR in squamous cell carcinoma, low-grade squamous intraepithelial lesion (LSIL) and high grade squamous intraepithelial lesions (HSIL) positive rate higher, the difference is statistically significant (P < 0. 05); compared with the wet wart and cervicitis, HC2 assay in squamous cell carcinoma (SCC), LSIL and HSIL positive rate higher, the difference is statistically significant (P < 0. 05), see table 1.
Table two comparison of 1 methods for the detection of high risk type HPV results (n,%)
2, multiplex real-time PCR detection of high-risk HPV subtype infection in the 120 cases of patients, the positive rate of PCR detection for 84. 2% (101/120), among them 26 cases for type 16, accounting for 25%; 21 cases for 18, accounting for 20%; 10 cases of 31 type, accounting for 9. 9%; 5 cases for 33, accounting for 5. 0%; 10 cases of 45 type, accounting for 9. 9%; 4 cases 52 type, accounting for 4. 0%. 4 cases were 56 type, accounting for 4. 0%; 21 cases 58 type, accounting for 20%. Type 18, type 16 and type 58 had a higher rate of infection, and the difference was statistically significant (P < 0.05).
3, discuss
Multiplex real-time PCR detection technology to a nucleic acid hybridization based, and is widely used to detect mRNA of gene technology, effectively reduce the polymerase chain reaction after the operation, also in different concentrations of mRNA compared with higher kinetic advantage [4]. Real time fluorescence quantitative PCR is the probe or dye fluorophores with PCR reaction system, the fluorescence signal accumulation to real-time monitoring of the whole process of PCR and DNA double strand infiltration of fluorescent dye a fluorescence signal, for both fluorescence signal and PCR product increased at the same time to provide effective protection, and fluorescent probe can in the probe of the 5 'end and the 3' end were labeled with a fluorescent reporter group, quenching quenching groups each, each other the role, the formation of structure of energy transfer. The quencher can effectively inhibit or absorption from the 5 'end of the fluorescence of the fluorophore; and once beyond a certain distance, the inhibitory effect disappeared. At this time the fluorescence signal increases, and then monitoring system to detect and receive [5].
By observing the intensity of the detected fluorescence signal, we can make an efficient detection of the gene product obtained after HPV amplification. The research results show that in the detection of high-risk HPV, this study used two methods for detecting efficiency approximation, there was no statistically significant differences (P > 0. 05); and compared with the wet wart and cervicitis, squamous cell carcinoma, low-grade squamous intraepithelial lesion (LSIL) and high grade squamous intraepithelial lesions (HSIL) positive rate higher, the difference is statistically significant (P < 0. 05).
To sum up, the multiplex real-time PCR method for the detection of easy automation, with good repeatability, detection is fast and simple, also has the sensitivity, specificity and high yield characteristics, DNA amplification product detection accuracy rate is high, suitable for monitoring high risk HPV persistent infection and large scale screening, clinical can be popularized and used.
reference
[1] Bing Yu Zou, Pu Haiying, Li Dan, et al. 210 cases of high-risk human papillomavirus papilloma virus infection detection and cervical lesions affecting factors study [J]. Journal of Practical Gynecology and obstetrics, 2013, 29 (12): 938-940.
[2] Hu Zhong Xiang, Li Dequn, Xi Cheng Yue, et. Breast cancer in high-risk human papilloma virus infection and with micro vascular density correlation [J]. Clinical and Experimental Pathology Journal, 2014, 30 (3): 261-263 (in Chinese with English Abstract).
Wei Wei, Song Yuntao, Zhang Baoyun, et al. Detection and analysis of human papilloma virus infection in patients with laryngeal squamous cell carcinoma (J). Journal of experimental and clinical research of China, 2013, 27 (1): 22-24.
Xing Zhiyan, Xu Dongyan, Hao Bai Lian, et al. Analysis of 955 cases of gynecological gynecological infections in women with HPV infection (J). Journal of reproductive medicine, 2013, 22 (12): 951-953.
[5] Shuang Shuang Nie, Ding Xian Ping, Chen Zuyi,. Chengdu region of human papilloma virus infection subtypes and age distribution, multiple infection and trend research [J]. International Journal of medical microbiology, 2013, 34 (22): 3026-3028.