Journal List > J Gynecol Oncol > v.28(5) > 1093837

Sun, Song, Ruan, Mao, Kang, Dong, and Lin: Clinical validation of the PCR-reverse dot blot human papillomavirus genotyping test in cervical lesions from Chinese women in the Fujian province: a hospital-based population study

Abstract

Objective

To determine the clinical significance of the polymerase chain reaction (PCR)-reverse dot blot (RDB) human papillomavirus (HPV) genotyping assay in cervical cancer screening.

Methods

A total of 10,442 women attending the Fujian Provincial Maternity and Children's Health Hospital were evaluated using the liquid-based cytology (thinprep cytologic test [TCT]) and the PCR-RDB HPV test. Women with HPV infection and/or abnormal cytology were referred for colposcopy and biopsy. For HPV DNA sequencing, 120 specimens were randomly selected. Pathological diagnosis was used as the gold standard.

Results

Using the PCR-RDB HPV test, overall HPV prevalence was 20.57% (2,148/10,442) and that of high-risk (HR)-HPV infection was 18.68% (1,951/10,442). There was 99.2% concordance between HPV PCR-RDB testing and sequencing. In this studied population, the most common HR-HPV types were HPV-16, −52, −58, −18, −53, −33, and −51, rank from high to low. HPV-16, −18, −58, −59, and −33 were the top 5 prevalent genotypes in cervical cancer but HPV-16, −18, −59, −45, and −33 were the top 5 highest risk factors for cancer (odds ratio [OR]=34.964, 7.278, 6.728, 6.101, and 3.658; all p<0.05, respectively). Among 10,442 cases, 1,278 had abnormal cytology results, of which, the HR-HPV positivity rate was 83.02% (1,061/1,278). To screen for cervical cancer by PCR-RDB HPV testing, when using CIN2+, CIN3+, and cancer as observed endpoints, the sensitivity was 90.43%, 92.61%, and 94.78% and the negative predictive value (NPV) was 99.06%, 99.42%, and 99.78%, respectively. PCR-RDB HPV and TCT co-testing achieved the highest sensitivity and NPV.

Conclusion

For cervical cancer screening, the PCR-RDB HPV test can provide a reliable and sensitive clinical reference.

References

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Fig. 1.
Prevalence of different HPV genotype infections. (A) HPV-16 is most prevalent genotype in HPV-positive women. (B) The HR-HPV infection rate showed a bimodal trend, and the rates of different age groups were statistically significant (χ2=272.740; p<0.001). Top 5 most frequent HPV genotypes in (C) NILM patients; (D) LSIL (CIN1) patients; (E) HSIL patients (CIN2 and CIN3); and (F) cervical cancer. Orange bar in (C-F) is the cumulative cases of HPV genotypes. CIN, cervical intraepithelial neoplasia; HPV, human papillomavirus; HR-HPV, high-risk human papillomavirus; HSIL, high-grade squamous intraepithelial lesion; LSIL, low-grade squamous intraepithelial lesion; NILM, negative for intraepithelial lesion or malignancy.
jgo-28-e50f1.tif
Fig. 2.
Age-related distribution of HPV infection. Top 5 most frequent HPV genotypes in different age groups of (A) NILM patients; (B) LSIL (CIN1) patients; (C) HSIL patients (CIN2 and CIN3); and (D) cervical cancer. CIN, cervical intraepithelial neoplasia; HPV, human papillomavirus; HSIL, high-grade squamous intraepithelial lesion; LSIL, low-grade squamous intraepithelial lesion; NILM, negative for intraepithelial lesion or malignancy. (continued to the next page)
jgo-28-e50f2.tif
Fig. 2.
(Continued) Age-related distribution of HPV infection. Top 5 most frequent HPV genotypes in different age groups of (A) NILM patients; (B) LSIL (CIN1) patients; (C) HSIL patients (CIN2 and CIN3); and (D) cervical cancer. CIN, cervical intraepithelial neoplasia; HPV, human papillomavirus; HSIL, high-grade squamous intraepithelial lesion; LSIL, low-grade squamous intraepithelial lesion; NILM, negative for intraepithelial lesion or malignancy.
jgo-28-e50f3.tif
Fig. 3.
Cumulative occurred risk of HR-HPV genotype for cervical lesions. Cumulative occurred risk of each HPV genotype: in patients with (A) a pathologically diagnosed CIN1; (B) CIN2; (C) CIN3; and (D) invasive cervical cancer. Orange bar represents statistically significant (p<0.05) green bar represents not statistically significant (p>0.05). CIN, cervical intraepithelial neoplasia; HPV, human papillomavirus; HR-HPV, high-risk human papillomavirus.
jgo-28-e50f4.tif
Table 1.
HPV infection by age related distribution (n=10,442)
Age (yr) Negative
(n=8,294)
Single-type HPV
(n=1,449)
Dual-type HPV*
(n=466)
Multi-type HPV
(n=233)
Total HPV infection
(n=2,148)
≤20 (n=89) 63 5 11 10 26 (29.21)
21–25 (n=861) 717 92 30 22 144 (16.72)
26–30 (n=1,749) 1,502 161 56 30 247 (14.12)
31–35 (n=1,693) 1,388 201 72 32 305 (18.02)
36–40 (n=1,765) 1,396 271 80 18 369 (20.91)
41–45 (n=1,853) 1,455 278 84 36 398 (21.48)
46–50 (n=1,335) 1,009 238 60 28 326 (24.42)
51–55 (n=537) 376 113 32 16 161 (29.98)
56–60 (n=284) 186 54 24 20 98 (34.51)
61–65 (n=164) 117 21 11 15 47 (28.66)
≥66 (n=112) 85 15 6 6 27 (24.11)

Values are presented as number (%). Detected by PCR-RDB genotyping, 18 HR-HPV, and 5 LR-HPV subtypes could be analyzed in the population. HPV, human papillomavirus; HR-HPV, high-risk human papillomavirus; LR-HPV, low-risk human papillomavirus; PCR-RDB, polymerase chain reaction-reverse dot blot.

* Dual-type infection: patients with 2 HPV subtypes infection simultaneously;

Multi-type infection: patients with 3 or more HPV subtypes infection simultaneously;

Represented the percent of HPV-positive patients in different age groups.

Table 2.
Compare HPV infection and TCT with pathological diagnosis (n=10,442)
Subjects Pathologic diagnosis χ2* p-value χ2 p-value
NILM (n=9,309) CIN1 (n=395) CIN2 (n=227) CIN3 (n=292) Cancer (n=219)
TCT (n=10,442)
HPV-positive (n=2,148) 1,155 333 197 262 201        
HPV-negative (n=8,294) 8,154 62 30 30 18        
Cytological diagnosis
NILM (n=9,164) 743.729 0.001 1,125.568 0.001
HPV-positive (n=1,087) 890 63 52 49 33
HPV-negative (n=8,077) 8,027 14 19 12 5
ASC (n=587) 22.125 0.001 38.252 0.001
HPV-positive (n=437) 168 142 41 57 29
HPV-negative (n=150) 99 36 6 6 3
LSIL (n=312) 0.584 0.540 3.103 0.524
HPV-positive (n=283) 68 118 47 39 11
HPV-negative (n=29) 6 11 4 5 3
≥HSIL (n=317) 10.688 0.011 10.741 0.020
HPV-positive (n=300) 10 4 53 111 122
HPV-negative (n=17) 4 0 1 7 5
≥AGC (n=62)§ 5.866 0.019 9.103 0.038
HPV-positive (n=41) 19 6 4 6 6
HPV-negative (n=21) 18 1 0 0 2

ASC, atypical squamous cells; AGC, atypical glandular cells; CIN, cervical intraepithelial neoplasia; HPV, human papillomavirus; HSIL, high-grade squamous intraepithelial lesion; LSIL, low-grade squamous intraepithelial lesion; NILM, negative for intraepithelial lesion or malignancy; TCT, thinprep cytologic test.

* χ2: the difference of patients with CIN2+ between the HPV-positive and -negative groups;

χ2: the difference between the HPV-positive and -negative groups in patients with different pathologic diagnosis and cytological diagnosis;

≥HSIL: HSIL and cervical squamous carcinoma;

§ ≥AGC: AGC and cervical adenocarcinoma.

Table 3.
Compare cytology and HPV in the different degree of cervical lesion detection (n=10,442)
Variable Sensitivity Specificity PPV NPV PLR NLR
CIN2+*
HPV 90.43 (88.21–92.65) 84.83 (84.12–85.55) 31.02 (29.05–32.98) 99.06 (98.85–99.27) 5.8977 (5.5919–6.2202) 0.1246 (0.1010–0.1537)
TCT 76.96 (73.93–80.00) 92.73 (92.21–93.25) 44.65 (41.92–47.39) 98.14 (97.86–98.42) 10.5837 (9.7569–11.4807) 0.2484 (0.2177–0.2835)
HPV+TCT 95.26 (93.72–96.79) 83.04 (82.29–83.79) 29.98 (28.12–31.83) 99.57 (99.42–99.71) 5.6162 (5.3588–5.8861) 0.0571 (0.0413–0.0789)
CIN3+
HPV 92.61 (89.08–93.64) 83.20 (82.46–83.93) 21.76 (20.00–23.51) 99.42 (99.26–99.58) 5.3918 (5.1188–5.6793) 0.1129 (0.0862–0.1478)
TCT 80.63 (77.20–84.05) 91.32 (90.77–91.87) 32.39 (29.82–34.96) 98.92 (98.71–99.13) 9.2889 (8.6029–10.0296) 0.2122 (0.1777–0.2532)
HPV+TCT 96.67 (95.12–98.23) 81.32 (80.55–82.09) 21.07 (19.75–22.72) 99.79 (99.69–99.89) 5.1747 (4.9514–5.4081) 0.0409 (0.0256–0.0653)
Cervical cancer
HPV 94.78 (91.14–95.42) 81.11 (80.35–81.87) 9.45 (8.27–10.69) 99.78 (99.68–99.88) 4.8581 (4.5915–5.1403) 0.1013 (0.0651–0.1578)
TCT 82.65 (77.63–87.66) 89.30 (88.70–89.90) 14.23 (12.31–16.15) 99.58 (99.45–99.72) 7.7270 (7.1142–8.3925) 0.1943 (0.1455–0.2594)
HPV+TCT 97.72 (95.74–99.70) 79.11 (78.31–79.90) 9.13 (8.65–10.29) 99.94 (99.88–99.99) 4.6772 (4.4810–4.8820) 0.0289 (0.0121–0.0686)

Values are presented as value (95% CI). CI, confidence interval; CIN, cervical intraepithelial neoplasia; HPV, human papillomavirus; NLR, negative likelihood ratio; NPV, negative predictive value; PLR, positive likelihood ratio; PPV, positive predictive value; TCT, thinprep cytologic test.

* CIN2+: included CIN2, CIN3, and cervical carcinoma;

CIN3+: included CIN3 and cervical cancer.

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