A new PCR-based mass spectrometry system for high-risk HPV, part I: methods.

Infection with high-risk (HR) human papillomaviruses (HPVs) has been confirmed as the necessary cause of cervical cancer. There are many studies that have established and confirmed the relationship of specific HPV types and the risk of invasive cervical cancer. We have developed a novel genotyping method for detecting 14 HR-HPV genotypes simultaneously with MassARRAY (Sequenom, San Diego, CA) technique based on the matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). All 14 HPVs showed high specificities and high sensitivities in the plasmid test; lower detection limits for each genotype were from 10 to 100 copies. Furthermore, the MS system has high-throughput capacities, capable of processing, with type-specific output, 4,500 samples in 24 hours. The MS HPV assay is a sensitive and useful tool for HPV genotyping. It has the potential to be suitable for large-scale epidemiologic studies and routine diagnostic clinical applications owing to its high-throughput capacity, high sensitivity, and low cost per case.

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.