| ObjectiveCurrently, cervical cancer is an only clear cause cancer. The main reason for cervical cancer is high-risk HPV persistent infection. The incidence of cervical cancer has become a major threat to women’s health killer. It was reported that every year there are more and more women suffering from cervical cancer. The resulting reduction of the quality of life as well as death has attracted the attention of the World Health Organization. Therefore, early diagnosis and prevention of cervical cancer is becoming more and more important.In some developed countries, the early screening of cervical cancer has spread. Pap smears greatly promote cervical cancer screening, however, the amount of its cells taken unstable, the diagnostic results affected by the impact of the cells can not be repeated, which limit its clinical diagnostic value. The birth of the liquid-based cytology has been radically improved the insufficient of cell. But it remain rely on the subjective judgment of the pathologist as a Pap smear. The diagnosis of pathologist, which reduces the sensitivity, is less constant. Moreover, abnormal cytology morphology occur relatively late, not early herald lesions, affecting its early diagnosis. Based on these factors, HPV detection becomes the entry point of early cervical cancer screening, which is also recognized by domestic and foreign counterparts.With the popularity of HPV HC2method, it becomes the gold standard of American FDA certification for detecting HPV DNA, because of the high sensitivity and stability. It has played an irreplaceable role in cervical cancer screening. However, with the deepening of research, the problem is gradually emerging. There are many reports that the specificity of detection is too low, the false positive rate is too high, this result leads clinicians take excessive treatment, which brought a lot of damage to the patient’s physical and psychological. The fundamental reason of low specificity is that HPV infection is universal and transient. HC2testing positive results can only prove who had HPV infection, whether the current body has cleared the virus or not infection continue can be confirmed. Even if the virus is still present in the body, and its integration into the host gene, it leading to disease, active replication and transcription, are unable to be read out from the experimental results of HC2. Thus HC2has its limitation.In this context, HPV E6/E7mRNA becomes a new biomarker, which can reflect the current body of virus activity and has greater clinical significance. The purpose of this study is to compare HPV E6/E7mRNA with the traditional classic method HC2from the sensitivity and specificity of diagnosis of cervical cancer, with histopathology as the gold standard.Materials and Methods1Clinical specimensThere are272cases cervical cytology specimens in our study, which is collected from January2011to December2011in Maternal and Child Health Hospital of Henan Province. These samples contain28cases of NILM,140cases of ASC-US,41cases of LSIL,47cases of HSIL,1case of SCC and15cases of AGC°The residual liquid specimens were collected for testing HPV E6/E7mRNA. All participants signed informed consent after told study details. 2Methods2.1HPV DNA testing by Hybrid Capture2(HC2)Using Digene dedicated HPV brush placed in the cervix, counter-clockwise three times for10seconds; put it in a dedicated specimen storage tube, repeatedly rinsed, broken part of the excess, cover the lid. The specimens were sent to the laboratory, five steps after lysis, hybridization, capture, antigen-antibody reaction and signal amplification, and finally placed in a dark place, read amplified signal by DML2000.2.2HPV E6/E7mRNA testing by Quantivirus? HPV E6/E7RNA3.0assay (bDNA)Using QUANTIVIRUS9HPV E6/E7RNA3.0Assay (bDNA) detection kit produced by DiaCarta, detecting HPV E6/E7mRNA of the remainder liquid-based cytology specimens. After rinsing, cracking, configuration, layout, signal amplification, plate reading, the results determined steps, we draw the HPV E6/E7mRNA detection results.2.3Cytopathological diagnosisCervical exfoliated cells were collected by the brush (Digene), which was washed deeply and repeatedly in preservation liquid to make the cells be washed down completely. Secondly, the ThinPrep?2000was used to make smears, then fix in95%alcohol. The smears, stained by Papanicolaou, mounted by neutral gum, were diagnosed blinded according to the TBS2001system by the three seniority of cytopathology physician under light microscope.2.4Histopathological diagnosisPatients with abnormal colposcopy were recommended to get a biopsy. The results of cervical biopsy were read by one or more academic pathologists and served as the disease endpoint for study purposes. All slides were diagnosed according to the current World Health Organization classification. Pathologists were blinded to HPV results.3Statistical analysisAll statistical analyses were performed using SPSS17.0statistical package for Windows. Pearson’s Χ2test was used to evaluate the significance of differences between designated groups. It was0.05for a to the significant level. ROC curve was used to assess the diagnostic accuracy (sensitivity and specificity) of Hybrid Capture2and E6/E7mRNA.Results1Cytological or histological follow-up diagnosis and positivity rate for HPV E6/E7messenger RNA test200out of272(73.5%) cases were mRNA positive.94out of111cases of histological diagnosis (84.9%) were mRNA positive.2Comparative performance of E6/E7mRNA and Hybrid Capture2assaysIn146patients with the E6/E7mRNA assay and the Hybrid Capture2assay, the consistency of the two methods was74.7%(109/146)(95%confidence interval [CI],67.6,81.8).3Correlation of E6/E7mRNA and HC2results with CIN2+There were75specimens simultaneous detection with four methods. The HPV mRNA assay and the HC2assay had the same sensitivity of high grade cervical intraepithelial neoplasia (CIN2+),82.4%(14/17)(95%confidence interval [CI],64.3,100). However, the specificity of CIN2+for the HPV mRNA and HC2assay was15.5%(95%confidence interval [CI],6.2,24.8) and29.3%(95%confidence interval [CI],17.6,41.0), which has no significant difference(P>0.05). 4ROC curveThe area under the curve of the HPV E6/E7mRNA and HC2methods were0.638and0.563, respectively. When the Youden index was maximum, the sensitivity and specificity of HPV E6/E7mRNA diagnostic were58.8%and74.1%, respectively. And the sensitivity and specificity of HPV HC2were47.1%and70.7%, respectively.Conclusion1The sensitivity of the HPV E6/E7mRNA assay was as same as HPV HC2(82.4%), the specificity has no significant difference with HC2.2HPV E6/E7mRNA can be used as a screening tool for cervical cancer.3HPV E6/E7mRNA can combine with HPV DNA to diagnose CIN2+better. |
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