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Development of a fluorescence-based multiplex genotyping method for simultaneous determination of human papillomavirus infections and viral loads.

Sun Z, Zhang R, Liu Z, Liu C, Li X, Zhou W, Yang L, Ruan Q, Zhang X - BMC Cancer (2015)

Bottom Line: In addition, the sensitivity and positive predictive value of BMRT is 98.4 % and 95.7 % compared with the results detected by PCR-sequencing method, respectively.With this cutoff value, 74.6 % sensitivity and 72.5 % specificity for prediction of HPV-16 infected patients with CINI and higher CIN were achieved.High significance was obtained when comparing the infected women presenting NILM/cervicitis with women either with CIN and cervical carcinomas (p < 0.001).

View Article: PubMed Central - PubMed

Affiliation: Virus Laboratory, The Affiliated Shengjing Hospital, China Medical University, Shenyang, Liaoning, 110004, China. sunzr@sj-hospital.org.

ABSTRACT

Background: Persistent high-risk human papillomavirus (HPV) infection is correlated with an increased risk of developing intraepithelial lesion or malignancy (NILM). The aims of the current study is to establish a method named BioPerfectus Multiplex Real Time (BMRT) HPV assay for simultaneous typing and quantifying HPVs, and to evaluate it by comparison with HPV GenoArray test and PCR-sequencing method, as well as histological status.

Methods: A total of 817 cervical specimens were evaluated by BMRT method and HPV GenoArray test, using PCR-sequencing method as the reference standard; simultaneously, high-risk HPV-16 and -18 DNA loads were assessed in 443 specimens to investigate the correlation with infection outcomes.

Results: The overall detection coincidence rate between BMRT assay and HPV GenoArray test is 96.6 % and the Kappa value is 0.760. In addition, the sensitivity and positive predictive value of BMRT is 98.4 % and 95.7 % compared with the results detected by PCR-sequencing method, respectively. HPV-16 viral load has a correlation with CINs or worse lesions. By comparing with infected women presenting NILM /cervicitis, the cutoff value for HPV-16 from patients with CINs was 0.827. With this cutoff value, 74.6 % sensitivity and 72.5 % specificity for prediction of HPV-16 infected patients with CINI and higher CIN were achieved. High significance was obtained when comparing the infected women presenting NILM/cervicitis with women either with CIN and cervical carcinomas (p < 0.001).

Conclusions: The BMRT assay seemed to be a good alternative approach for HR-HPV testing, due to its high level of automation and ability to quantify HPV-16, HPV-18 and other HR-HPVs.

No MeSH data available.


Related in: MedlinePlus

Comparsion of HPV - 18 viral loads between NILM/cervicitis and ASCUS/CIN I-III/cancer
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Fig4: Comparsion of HPV - 18 viral loads between NILM/cervicitis and ASCUS/CIN I-III/cancer

Mentions: Among 130 HPV-18 positive patients, 81 were negative in histological diagnosis (including normal, infusorian and cervicitis), 27 were LSIL/ASCUS/ASC-H/CINI, 14 were HSIL/CINII-III, and 8 were cervical cancer. The relative levels of HPV-18 DNA loads (copies per 10,000 cells) in corresponding cervical scrapes were calculated to correlate them with the histopathological findings of infected patients (Fig. 3). The median viral load (lg) in patients with diagnosis of NILM/cervicitis, LSIL/ASCUS/ASC-H/CINI, HSIL/CINII-III and cancer were 3.21, 4.84, 3.83 and 3.92, respectively. Moreover, pairwise comparisons among the four groups by Kruskal-Wallis test showed highly significant difference of viral loads in samples between the group presenting NILM/cervicitis and the group of LSIL/ASCUS/ASC-H/CINI (p < 0.001). Unlike those of HPV16, the viral loads of HPV-18 were less associated with progress of CINII-III. Similar to those of HPV-16, the significant differences of HPV-18 viral loads were observed when comparing the group presenting NILM/cervicitis to either all grades of CIN (p < 0.001), or to cervical carcinomas group (p < 0.001) by Mann–Whitney U test. As shown in Fig. 4, the median viral loads (copies per 10,000 cells) in patients with NILM/cervicitis and ASCUS/CINI-III/cancer were 3.21 and 4.31, respectively.Fig. 3


Development of a fluorescence-based multiplex genotyping method for simultaneous determination of human papillomavirus infections and viral loads.

Sun Z, Zhang R, Liu Z, Liu C, Li X, Zhou W, Yang L, Ruan Q, Zhang X - BMC Cancer (2015)

Comparsion of HPV - 18 viral loads between NILM/cervicitis and ASCUS/CIN I-III/cancer
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4635533&req=5

Fig4: Comparsion of HPV - 18 viral loads between NILM/cervicitis and ASCUS/CIN I-III/cancer
Mentions: Among 130 HPV-18 positive patients, 81 were negative in histological diagnosis (including normal, infusorian and cervicitis), 27 were LSIL/ASCUS/ASC-H/CINI, 14 were HSIL/CINII-III, and 8 were cervical cancer. The relative levels of HPV-18 DNA loads (copies per 10,000 cells) in corresponding cervical scrapes were calculated to correlate them with the histopathological findings of infected patients (Fig. 3). The median viral load (lg) in patients with diagnosis of NILM/cervicitis, LSIL/ASCUS/ASC-H/CINI, HSIL/CINII-III and cancer were 3.21, 4.84, 3.83 and 3.92, respectively. Moreover, pairwise comparisons among the four groups by Kruskal-Wallis test showed highly significant difference of viral loads in samples between the group presenting NILM/cervicitis and the group of LSIL/ASCUS/ASC-H/CINI (p < 0.001). Unlike those of HPV16, the viral loads of HPV-18 were less associated with progress of CINII-III. Similar to those of HPV-16, the significant differences of HPV-18 viral loads were observed when comparing the group presenting NILM/cervicitis to either all grades of CIN (p < 0.001), or to cervical carcinomas group (p < 0.001) by Mann–Whitney U test. As shown in Fig. 4, the median viral loads (copies per 10,000 cells) in patients with NILM/cervicitis and ASCUS/CINI-III/cancer were 3.21 and 4.31, respectively.Fig. 3

Bottom Line: In addition, the sensitivity and positive predictive value of BMRT is 98.4 % and 95.7 % compared with the results detected by PCR-sequencing method, respectively.With this cutoff value, 74.6 % sensitivity and 72.5 % specificity for prediction of HPV-16 infected patients with CINI and higher CIN were achieved.High significance was obtained when comparing the infected women presenting NILM/cervicitis with women either with CIN and cervical carcinomas (p < 0.001).

View Article: PubMed Central - PubMed

Affiliation: Virus Laboratory, The Affiliated Shengjing Hospital, China Medical University, Shenyang, Liaoning, 110004, China. sunzr@sj-hospital.org.

ABSTRACT

Background: Persistent high-risk human papillomavirus (HPV) infection is correlated with an increased risk of developing intraepithelial lesion or malignancy (NILM). The aims of the current study is to establish a method named BioPerfectus Multiplex Real Time (BMRT) HPV assay for simultaneous typing and quantifying HPVs, and to evaluate it by comparison with HPV GenoArray test and PCR-sequencing method, as well as histological status.

Methods: A total of 817 cervical specimens were evaluated by BMRT method and HPV GenoArray test, using PCR-sequencing method as the reference standard; simultaneously, high-risk HPV-16 and -18 DNA loads were assessed in 443 specimens to investigate the correlation with infection outcomes.

Results: The overall detection coincidence rate between BMRT assay and HPV GenoArray test is 96.6 % and the Kappa value is 0.760. In addition, the sensitivity and positive predictive value of BMRT is 98.4 % and 95.7 % compared with the results detected by PCR-sequencing method, respectively. HPV-16 viral load has a correlation with CINs or worse lesions. By comparing with infected women presenting NILM /cervicitis, the cutoff value for HPV-16 from patients with CINs was 0.827. With this cutoff value, 74.6 % sensitivity and 72.5 % specificity for prediction of HPV-16 infected patients with CINI and higher CIN were achieved. High significance was obtained when comparing the infected women presenting NILM/cervicitis with women either with CIN and cervical carcinomas (p < 0.001).

Conclusions: The BMRT assay seemed to be a good alternative approach for HR-HPV testing, due to its high level of automation and ability to quantify HPV-16, HPV-18 and other HR-HPVs.

No MeSH data available.


Related in: MedlinePlus