SNIPER: a novel assay for human papillomavirus testing among women in Guizhou, China.

OBJECTIVE: Clinically validate the SNIPER human papillomavirus (HPV) DNA assay for the detection of cervical intraepithelial neoplasia (CIN)2 or higher and CIN2 or higher in a prospective cross-sectional screening study in Guizhou Province, China. METHODS: Between March and April, 2008, 1000 nonpregnant women aged 30 or older were recruited in Guizhou Province, China. Women positive by SNIPER or cytological examination were requested to return for follow-up. A biopsy of all colposcopically detected abnormalities was performed by quadrant. In normal quadrants, biopsies were obtained at the squamocolumnar junction (2-, 4-, 8-, and 10-o'clock positions depending on the quadrant). Samples were placed in 2 mL of saline solution and maintained between 2 degrees C and 30 degrees C for up to 1 week. One milliliter of this suspension was then prepared and tested. For polymerase chain reaction amplification, a pool of HPV primers was designed to amplify HPV DNA from 13 high-risk-HPV genotypes (types 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, and 68). Test characteristics were calculated according to standard definitions. RESULTS: One thousand women were screened; 175 tested HPV positive, 36 women tested negative but had positive Papanicolaou test results. All but 21 (90%) returned for follow-up. Median age and proportions having CIN2 or higher and CIN3 or higher differed by HPV status. Twenty-five women had CIN2 or higher and 16 had CIN3 or higher. The SNIPER assay was 93.3% and 94% sensitive and 86% and 85% specific for the detection of CIN2 or higher and CIN3 or higher, respectively. The positive predictive value was 17.4 % and 9.9% for CIN2 or higher and CIN3 or higher, respectively. Negative predictive value approached 100% for CIN2 or higher and CIN3 or higher. CONCLUSION: The SNIPER assay is functionally competitive and in terms of cost holds an advantage over Hybrid Capture 2 in a Chinese healthcare market, and potentially others, around the world.

Study of the diagnostic efficacy of real-time optical coherence tomography as an adjunct to unaided visual inspection with acetic acid for the diagnosis of preinvasive and invasive neoplasia of the uterine cervix.

OBJECTIVES: To determine the sensitivity and specificity of optical coherence tomography (OCT) as an adjunct to unaided visual inspection using acetic acid (VIA) in the detection of cervical intraepithelial neoplasia 2 (CIN 2) in a real-time clinical evaluation. BACKGROUND: This clinical study was a prospective cross-sectional comparative trial that screened 1000 patients (aged 30-50 years) in a low-resource setting. Women with abnormal cervical cytology or positive human papillomavirus (HPV) tests were referred for further evaluation including VIA, OCT imaging, colposcopy, and cervical biopsies. METHODS: The VIA diagnoses were coded by quadrant. The OCT was then performed in all VIA-positive areas and at the squamocolumnar junction in all 4 quadrants. All patients were colposcoped; assessed by quadrant with biopsies at 2, 4, 8, and 10 o'clock; all abnormal areas were biopsied; and endocervical curettage was performed. Data were analyzed using generalized estimating equations and logistic regression. RESULTS: Of the 1000 patients, 175 (17.5%) were HPV positive, 93 (9.3%) had abnormal cervical cytology greater than or equal to atypical squamous cells of undetermined significance, and 211 (21.1%) were either HPV positive or had abnormal cytology. The VIA, OCT, colposcopy, and biopsies were completed on 183 (86.7%) of 211 women. For VIA alone, the sensitivity and specificity in detecting lesions greater than or equal to CIN 2 was 43% and 96%. With the addition of OCT, the sensitivity increases to 62% with a specificity of 80%. CONCLUSIONS: With the addition of OCT, the sensitivity of VIA increased in all analyses for the detection of greater than or equal to CIN II, with a loss in specificity. We hope that the potential of this technology will be realized when a computer algorithm is generated to aid in image interpretation.

[Prevalence of high-risk human papillomavirus and incidence of cervical intraepithelial neoplasia in female populations in Shenzhen, Guangdong Province].

OBJECTIVE: To investigate the prevalence of high-risk human papillomavirus (HPV) and incidence of cervical intraepithelial neoplasia (CIN) in female populations in Shenzhen, Guangdong Province, China. METHODS: Totally 1137 women aged 15-59 from Shahe Community, Nanshan District, Shenzhen were investigated for cervical cancer during an population-based epidemiological screening from November 2004 to December 2004. Visual inspection with acetic acid (VIA), colposcopy, liquid-based cytology test (LCT), and hybrid capture 2 (HC-) were performed to detect the high-risk HPV types in cervical secretions. Biopsy under colposcope was performed in women who were HPV-positive with LCT >or= atypical squamous cells of undetermined sign (ASCUS) or HPV-negative with LCT >or= low grade squamous intraepithelial lesion (LSIL), with the pathological results as the golden standards. RESULTS: The detection rate of high-risk HPV-DNA was 14.0%. HPV detection rates in 15-24, 25-29, 30-34, 35-39, 40-44, 45-49, and 50-59 age groups were 15.5%, 17.7%, 12.6%, 8.8%, 10.2%, 15.3%, and 21.0%, respectively (P < 0.05). HPV detection rates in 25-29 years group and 50-59 years group were significantly higher than those in other groups (P < 0.05) and 35-39 group had the lowest detection rate. The curve of HPV infection rates in all groups was 'V' type. The overall incidence of CIN was 4.4%. The incidences of CIN , CIN , and CIN were 3.2%, 1.0%, and 0.3%, respectively, in which the incidence of CIN was significantly higher than those of CIN and . HPV detection rates increased with cervical lesion grades, which in >or=CIN groups and normal group were 100.0% and 8.3%, respectively. No cervical cancer was identified in this research. The sensitivities of VIA, colposcopy, LCT, and HC-II for high-risk HPV screening were 35.7%, 50.0%, 92.9%,and 100%, respectively, in detecting high-grade squamous intraepithelial lesion (HSIL), the specificities of these four methods were 96.0%, 87.2%, 88.4%, and 86.9%, respectively. Satisfactory negative predictive values were obtained for all methods. CONCLUSIONS: HPV infection is the main risk factor for CIN. Cervical cancer among female populations in Shenzhen is still in early stages. Prevention of HPV infection and treatment of CIN are key for the prevention of cervical cancer.

Diagnostic efficacy of real-time optical coherence tomography in the management of preinvasive and invasive neoplasia of the uterine cervix.

OBJECTIVE: Determine the sensitivity and specificity of optical coherence tomography (OCT) as an adjunct to colposcopy in the detection of cervical intraepithelial neoplasia (CIN) grade 2 or higher in a real-time clinical evaluation. BACKGROUND: Optical coherence tomography (OCT) uses infrared light similar to ultrasound pulse-echo imaging. Image resolution is optimal in the 1-to-3-mm range. This study is the third in our series of OCT investigations and our first real-time clinical trial. The study was conducted at the Peking University Shenzhen Hospital, Shenzhen, China. METHODS: Nonpregnant women 18 years or older with abnormal cervical cytologic findings or a positive high-risk human papillomavirus test result were recruited. Women were assessed; and diagnoses, recorded by cervical quadrant first with colposcopy, followed by colposcopic directed OCT. A biopsy of the abnormal areas was performed. In normal quadrants, biopsy specimens were obtained at the 2-, 4-, 8-, and 10-o'clock positions at the squamocolumnar junction depending on the quadrant. An endocervical curettage was also done. Individual OCT diagnoses were paired with colposcopic impressions and biopsy specimens to assess its role as a paired secondary screen. Data were analyzed using generalized estimating equations to control for correlation within a woman. RESULTS: One thousand two hundred thirty-seven paired diagnoses from 299 women were analyzed. Median age was 36 years. Ninety-six women (8%) had a diagnosis of CIN 2 or higher. Evaluation by quadrant showed that the sensitivity for CIN 2 or higher decreased by adding OCT to colposcopy, but the specificity increased from 83% to 93%. CONCLUSIONS: We continue to try to improve sensitivity by improving the near-infrared light source, decreasing the scan time to 8 frames per second, and using a larger diameter (5 mm) fiberoptic probe with a newly designed application specific probe sheath.

Genomic amplification of the human telomerase RNA gene for differential diagnosis of cervical disorders.

To evaluate genomic amplification of the human telomerase RNA gene (TERC) as a supportive approach to cytopathology or histopathology in diagnosis of low-grade and high-grade uterine cervical lesions, 1,033 Chinese women at three medical centers had liquid-based thin-layer cytopathologic examination and TERC detection by fluorescence in situ hybridization (FISH). Human papillomavirus DNA testing, colposcopy with or without biopsy, and histopathologic examination were conducted as needed. In cytopathologic examination, genomic amplification of TERC was found in 30 of 659 (4.6%) normal or benign cellular changes; in 23 of 170 (13.5%) atypical squamous cells of undetermined significance (ASCUS); in 8 of 28 (28.6%) atypical squamous cells with high-grade squamous intraepithelial lesion possible (ASC-H); and in 26 of 103 (25.2%) low-grade (LSIL) and 64 of 73 (87.7%) high-grade (HSIL) squamous intraepithelial lesions; with pairwise significant difference (P< 0.05) in each, except ASC-H and LSIL (chi(2) = 0.127, P = 0.72). In histopathologic examination, TERC was amplified in 28 of 671 (4.2%) normal, inflammatory, or wart cases; in 17 of 233 (7.3%) cervical intraepithelial neoplasia grade 1 cases (CIN 1); in 27 of 39 (69.2%) CIN 2 cases; in 57 of 67 (85.1%) CIN 3 cases; and in 22 of 23 (95.7%) cervical cancer cases; with pairwise significant difference in each (P < 0.05). The number of cells with abnormal signals increased and the abnormal signal patterns were diversified with increasing severity of cervical dysplasia. FISH detection of TERC amplification may provide an effective, noninvasive approach in conjunction with cytopathologic or histopathologic evaluation for differential diagnosis of low- and high-grade cervical disorders.

[Amplification and clinical significance of hTERC gene in the cervical exfoliated cells from natural population in Shenzhen].

OBJECTIVE: To study the amplification of human telomerase RNA component (hTERC) gene in the cervical exfoliated cells from natural population in Shenzhen and to explore its relationship with human pappiloma-virus (HPV) infection, abnormal cervical cytology and cervical intraepithelial neoplasia (CIN). METHODS: Three hundred and eighty-eight women, 30 - 59 year-old and having normal intelligence in a community of Shenzhen, were examined with liquid-based cytology. Human pappiloma-virus (HR-HPV) was tested by hybrid capture II (HC-II) and hTERC gene detection by fluorescence in situ hybridization (FISH). Patients with ASCUS and above lesion, and/or positive HR-HPV results and/or amplification of hTERC gene were examined by colposcopy, multiple biopsies of cervical quadrant and pathology. RESULTS: The frequencies of CIN I, II, III, cervical cancer were 55 (14.18%), 4 (1.03%), 9 (2.32%) and 1 (0.26%) respectively, with the rate of hTERC gene amplification as 8.76%. There were significantly differences of hTERC amplification among the samples with different cytological and histological lesions as well with HPV infection (P < 0.01). (1) The positive rate of HPV infection was 17.01%; the positive rates of hTERC gene amplification were 19.70% in HPV positive and 6.52% in HPV negative samples and the results were significantly different (P < 0.01). (2) Cytologically, the rates of hTERC gene amplification appeared to be as follows: NILM (5.97%), ASCUS (18.75%), LSIL (10.00%), ASC-H (66.67%), HSIL (100.00%). There was a marked increase of hTERC amplification in patients with HSIL and above lesions (P < 0.01). (3) On histology findings, the rates of hTERC gene amplification were as follows: NILM (0%), CIN I (5.45%), CIN II (50.00%), CIN III (77.78%), and invasive carcinoma (100.00%). There was a marked increase of hTERC amplification in patients with CIN II and above lesions (P < 0.01). CONCLUSION: There was a close correlation between amplification of hTERC and histological as well cytological lesions which increased progressively along with the severity of cytological and histological grade. The evidence of hTERC, with or without amplification, might serve as a prognostic indicator to measure the grade of lesion.

[Clinical application of telomerase RNA component gene amplification assay in cervical lesions.].

OBJECTIVE: To investigate the significance of genomic amplification of the telomerase RNA component (TERC) gene to serve as a genetic biomarker in the screening of cervical lesions. METHODS: A total of 715 cases were recruited, with liquid-based cytology diagnosis as normal (n = 347), atypical squamous cells of undetermined significance (ASCUS, n = 180), atypical squamous cells cannot exclude a high-grade lesion (ASC-H, n = 13), low-grade squamous intraepithelial lesions (LSIL, n = 115), high-grade squamous intraepithelial lesions (HSIL, n = 59) and atypical glandular cells (AGC, n = 1). The remaining cervical cells in the cytological preserving fluid were analyzed using a two-color fluorescence in situ hybridization (FISH) probe targeted to chromosome 3q26 containing TERC gene. The TERC gene findings were compared to the cytological and histological detected results, as well as high-risk human papillomavirus (HPV) detected results. RESULTS: Genomic amplification of TERC gene was found in 5.8% of normal specimens, 22.2% of ASCUS, 30.8% of ASC-H, 27.8% of LSIL, 86.4% of HSIL and 1/1 of AGC. The positive rate was significantly lower in normal, ASCUS, ASC-H and LSIL compared with HSIL (all P < 0.01). Significantly more cells with genomic amplification of TERC gene were found in cervical intraepithelial lesion (CIN) II-III than CINI (77.8% vs. 9.3%), as well as invasive cervical cancer (96.7% vs. 9.3%), both P < 0.01. The rate of TERC gene amplification was higher in HPV positive patients (33.5%) than in HPV negative patients (5.2%, P < 0.01). The sensitivity of TERC gene amplification was significantly higher than that of cytological screening (81.88% vs. 36.96%, P < 0.01) in the differentiation of CINII or higher and CINI or lower diseases, its specificity was higher than high-risk HPV test (93.32% vs. 33.93%, P < 0.01) and positive prediction value (81.29%) was similar with cytological method (86.44%, P > 0.05); but its negative prediction value (93.56%) was lower than HPV test (97.06%, P < 0.05). CONCLUSIONS: The positive rates of TERC gene amplification increased as cervical diseases worsened. TERC gene amplification is related to HPV infection. The gain of chromosome 3q26 in cytological specimens is an effective molecular genetic biomarker in screening of CINII or higher and invasive cervical cancer.