The Performance Of Human Papillomavirus DNA Detection With Type 16/18 Genotyping By Hybrid Capture In Cervical Cancer Screening

Cervical cancer is the second most common cancer in developing countries and the third leading cause of cancer death.In China,cervical cancer accounts for the second highest incidence of gynecological cancer.Cervical cancer shows an increasing trend in morbidity,mortality,and the younger age of onset.Traditional cytology tests(Pap smear,liquid-based cytology)play an important role in cervical cancer screening,but their low senstivity,poor reproducibility and excessive medical resources requirement are also significant.High-risk HPV(HR-HPV)persistent infection is a necessary condition for the onset of cervical cancer.A number of HPV detection technologies have been developed and applied to cervical cancer screening.In 2006,the American Society of Colposcopy and Cervical Pathology(ASCCP)published a guideline for the recommendation of HPV testing combined with cytology screening as a screening strategy for cervical cancer in women over 30 years old.With the developed of HPV detection research in cervical cancer screening,molecular detection has gradually replayced of cytology detection in cervical cancer screening.The European Reproductive Organ Infection and Cancer Research Group(EUROGIN)recommended high-risk HPV testing for screening of cervical cancer in women over 25 years old as early as 2008.Epidemiology evidences confirmed the difference in pathogenicity of 14 high-risk types of HPV,among which HPV 16 and HPV 18 are most closely related to cervical cancer.Based on the important role of HPV16/18 in the development of cervical cancer,EUROGIN in 2010 proposed the importance of clinical application of HPV typing,HPV DNA detection with 16/18 genotyping techniques have also been developed and applied in clinic.Cobas 4800 HPV(Roche,USA)is a polymerase chain reaction(PCR)based HPV DNA assay and is the first FDA-approved HPV assay with HPV 16/18 typing.A large-scale clinical study of Cobas HPV in cervical cancer screening revealed that:HPV 16/18 typing showed a similar sensitivity and positive predictive value for cytology screening,greatly reducing reliance on cytologists,showing the prospect of good promotion of technology in areas with scarce medical resources.However,as a PCR-based technology,both real-time PCR and selective amplification in Cobas HPV assays require specific laboratories and qualified laboratory operators,which limit their widespread use in under-resourced area.DH3 HPV detection is a new technology based on hybrid capture principle that can detect 14 high-risk HPV DNA by HPV 16/18 genotyping.DH3 HPV Detection HPV only amplifies chemical signals without nucleic acid amplification,which allows the assay to be performed in a common laboratory with no special requirements and is suitable for cervical cancer screening in developing countries.However,by now,there have been no clinical trials from a large sample of the general population with a colposcopy biopsy as the gold standard to verify the performance of the DH3 HPV test reagent in predicting high-grade cervical lesions.This study selected women aged 21-65 years in some rural,urban and enterprise areas in Longyou County,Zhejiang Province to perform Cobas4800 HPV and cytology cotesting screening,residual cervical cytology samples were used for DH3 HPV testing.Taking colposcopy and/or Histopathological diagnosis as the standard,the clinical validation of the two HPV tests in cervical cancer screening were compared.Part I Comparion the efficacies between DH3 HPV and Cobas HPV for predicting high-grade cervical lesion Objectives:To compare the screening efficacy of two HPV testing with HPV 16/18 genotyping,PCR-based HPV testing(Cobas 4800,Roche)and hybrid capture-based HPV testing(DH3,Hangzhou Detong)in primary test of cervical cancer screening.Methods:Between April and May of 2015,332 administrative villages and communities were randomly selected from Longyou County in Zhejiang Province,China,and a program of cervical cancer screening was carried out using HPV testing(Cobas 4800,Roche)and cytology test(Thinprep,Hologic).Totally 11,356 women aged 21-65 years participated in this program.Residual samples were tested for DH3 HPV testing(Hangzhou Dalton Biosciences,Hangzhou,China).All women with positive Cobas HPV or abnormal cytology(ASC-US or worse)referred to colposcopy with or without biopsy;meanwhile 4.5%of women with both negative tests were randomly selected and referred to colposcopy.The study compared the performances between two HPV tests when they were respectively used in the strategy of primary HPV detection for cervical cancer screening in women aged 25 years and older.The effectiveness of detecting high-grade cervical lesions(including CIN2+ and CIN3+)was assessed using the sensitivity,specificity,positive predictive value(PPV),negative predictive value(NPV),positive likelihood ratio(PLR),and negative likelihood ratio(NLR),respectively.A non-inferiority analysis was used to determine whether the performance of DH3 HPV was equivalent to that of Cobas HPV.Result:1 A total of 10,669 women with valid Cobas HPV tests,DH3 HPV tests,cytology tests and colposcopy/pathology results were enrolled in the study.Among these women,420 women were diagnosed with CIN1,52 women were diagnosed with CIN2,83 women were diagnosed with CIN3+.Among 10669 women,1056(9.9%)women were DH3 HPV positive with 255(2.4%)16/18 positive,while 990(9.3%)were Cobas HPV positive with 243(2.3%)16/18 positive.2.The positive rates of TCT test were 24.5%,56.4%,59.6%and 83.1%,respectively in different pathological groups as inflammation or normal,CIN1,CIN2 and CIN3+,DH3 HPV positive rates were 32.5%,65%,86.5%and 97.6%,respectively;The positive rates of Cobas HPV were 47.5%,76.7%,88.5%and 92.8%,respectively.The positive rates of DH3 HPV 16/18 were 6.1%,14%,34.6%and 61.4%,respectively.The positive rates of Cobas HPV 16/18 were 8.7%,17.9%,36.5%and 59%,respectively.The positive rates of other types of DH3 HPV were 26.4%,51%,51.9%and 36.1%,respectively.The positive rates of other types of Cobas HPV were 38.8%,58.8%,51.9%and 33.7%.The positive rate of TCT and HPV detection increased with the severity of the lesion and the HPV 16/18 positive rate increased significantly with the severity of the lesion(X2= 273.27,DH3 HPV P<0.001,X2= 233.95,P<0.001)for Cobas HPV).3.Non-inferiority score test was used to compare the sensitivities between DH3 HPV and Cobas HPV for the predetermined threshold for CIN2+ and the specificities between two HPV tests for the predetermined threshold for CIN1-,and the results showed that DH3 HPV was non-inferior to Cobas HPV as primary HPV testing,among women aged 25 years and older(T=3.2,P=0.0008 and T=5.2,P<0.0001).Furthermore,the sensitivity,specificity,PPV,and NPV of DH3 HPV for identifying CIN2+ were 93.3%(95%CI:87.7-96.9),91.2%(95%CI:90.6-91.7),12.0%(95%CI:10.1-14.1),and 99.9%(95%CI:99.8-100),respectively,and all of them were not significantly different from those of Cobas HPV:sensitivity 91.1%(95%CI:85.0-95.3),specificity 91.8%(95%CI:91.2-92.3),PPV 12.5%(95%CI:10.5-14.7),and NPV 99.9%(95%CI:99.8-99.9),respectively(P>0.05).The similar results were observed for identifying CIN3+:the sensitivity,specificity,PPV,and NPV of DH3 HPV for identifying CIN3+ were 97.6%(95%CI:91.6-99.7)?90.8%(95%CI:90.2-91.4)?7.7%(95%CI:6.2-9.5)and 100%(95%CI:99.9-100),respectively,and all of them were not significantly different from those of Cobas HPV:sensitivity 92.8%(95%CI:84.9-97.3),specificity 91.4%(95%CI:90.8-91.9),PPV7.8%(95%CI:6.2-9.7),and 99.9%(95%CI:99.9-100.0),respectively(P>0.05).Conclusion:1.The positive rate of HPV 16/18 is significantly increased with the severity of cervical lesions,suggesting that HPV16/18 genotyping is important for the triage of high-risk HPV-positive people.2.DH3 HPV test with 16/18 genotyping shows no-inferior performance for identifying cervical high-grade lesions,compared with Cobas HPV test,and can be one of optional tests in cervical cancer screening for women aged 25 years older.Part ? Comparison the efficacies between HPV primary screening with 16/18 genotyping screening stratergis using DH3 HPV and Cobas HPVObjectives:To compare the effectiveness of hybrid capture-based HPV16/18 typing testing(DH3 HPV,Hangzhou Detong)and Cobas HPV(Cobas 4800,Roche)testing in primary HPV testing with HPV 16/18 genotyping,in which HPV16/18 positive refers to colposcopy and 12 other high-risk HPV positive refers to cytology triage.To certify the implementing of DH3 HPV test in cervical cancer screening programs in China.Methods:The research population and testing methods are the same as the first part.The study compared the performances between two HPV tests when they were respectively used in the strategy of primary HPV detection with 16/18 genotyping for cervical cancer screening in women aged 25 years and older.The effectiveness of detecting high-grade cervical lesions(including CIN2+ and CIN3+)was assessed using the sensitivity,specificity,positive predictive value(PPV),negative predictive value(NPV),positive likelihood ratio(PLR),and negative likelihood ratio(NLR),respectively.A non-inferiority analysis was used to determine whether the performance of DH3 HPV was equivalent to that of Cobas HPV.Result:1.Taking CIN2+ lesions as test endpoint,non-inferiority score test was used to compare the sensitivities between DH3 HPV and Cobas HPV for the predetermined threshold for CIN2+ and the specificities between two HPV tests for the predetermined threshold for CIN1-,and the results showed that DH3 HPV was non-inferior to Cobas HPV in primary HPV testing with HPV 16/18 genotyping screening strategy screening in women aged 25 years and older who received(T=3.6,P=0.0001 and T=10.0,P<0.0001).2.The sensitivity,specificity,positive predictive value and negative predictive value of CIN2+ detected by DH3 HPV test in 25 years and older women were 84.4%(95%CI:77.2-90.1),96.4%(95%CI:96.1-96.8),23.4%(95%CI:19.7-27.4)and 99.8%(95%CI:99.7-99.9)respectively.The sensitivity,specificity,positive predictive value and negative predictive value of Cobas HPV test were 80.0%(95%CI:72.3-86.4),96.7%(95%CI:96.4-97.1),24.0%(95%CI:20.1-28.2)and 99.7(95%CI:99.6-99.8)respectively.None of them showed significant difference(P>0.05);the sensitivity,specificity,positive predictive value and negative predictive value of CIN3+ detected by DH3 HPV test in 25 years and older women were 94.0%(95%CI:86.5-98.0),96.1%(95%CI:95.7-96.5),16.0%(95%CI:12.8-19.5)and 100.0%(95%CI:99.9-100.0),respectively;sensitivity,specificity,positive predictive value and negative predictive value of CIN3+ detected by Cobas HPV were 88.0%(95%CI:79.0-94.0),96.4%(95%CI:96.1-96.8),16.2%(95%CI:12.9-20.0)and 99.9%(95%CI:99.8-100.0),respectively.None of them showed significant difference(P>0.05).Conclusion:1.DH3 HPV test with 16/18 genotyping shows no-inferior performance with Cobas HPV test in the strategy of primary HPV detection with 16/18 genotyping for cervical cancer screening in women aged 25 years and older.2.Since DH3 HPV test is based on hybrid capture technology,which is simple,and inexpensive,low laboratory conditions and techenological level requirements,it seems more suitable for developing countries than Cobas HPV test does.