Comparative evaluation of AMPLICOR HPV PCR and Linear Array assays on SurePath liquid-based Pap samples for the detection of high-risk HPV genotypes

• Published in: Journal of clinical virology 2011-03, Vol.50 (3), p.201-204
• Main Authors: Chernesky, Max, Jang, Dan, Portillo, Eder, Smieja, Marek, Chong, Sylvia, Buracond, Sonya, Klingel, Michelle, Sebai, Rami El, Kapala, Julius, Patel, Jay, Biers, Kathy, Harroun, Nancy, Doucette, Chris, Sumner, Jeff
• Format: Article
• Language: English
• Subjects: Adult; Allergy and Immunology; AMPLICOR; Biological and medical sciences; DNA, Viral - genetics; Female; Fundamental and applied biological sciences. Psychology; Genotype; HC2; HPV; Human viral diseases; Humans; Infectious Disease; Infectious diseases; L-Pap; Linear array; Medical sciences; Microbiology; Middle Aged; Miscellaneous; Molecular Diagnostic Techniques - methods; Nucleic Acid Hybridization - methods; Papillomaviridae - classification; Papillomaviridae - genetics; Papillomaviridae - isolation & purification; Papillomavirus Infections - diagnosis; Papillomavirus Infections - virology; Polymerase Chain Reaction - methods; SurePath; Viral diseases; Virology; Virology - methods
• ISSN: 1386-6532; 1873-5967
• DOI: 10.1016/j.jcv.2010.12.003

Abstract Background Persistent cervical infection with high-risk [HR] HPV is a causative factor for cancer. Liquid-based [L-Pap] Pap samples are convenient for HPV testing and SurePath samples have been least studied. Most HPV tests have multiple step protocols and testing laboratories experience large volumes of samples. Objectives Using SurePath L-Pap residual samples the objectives were as follows: [1] to test the performance of AMP-HPV and LA-HPV. [2] To perform an agreement study between two laboratories for the AMP-HPV test and [3] to compare agreement of results between AMP-HPV and LA-HPV and HC2. Study design Samples from 657 women were tested for Pap cytology then assayed for HR-HPV using AMP-HPV and LA-HPV tests. AMP-HPV performance was compared between 2 laboratories and agreement studies were conducted between AMP-HPV, LA-HPV and HC2. Results HR-HPV genotypes were associated with L-Pap readings as follows: HSIL 92% [23/25], LSIL 73.6% [162/220], ASCUS 70.4% [131/186], normal 31.9% [72/226]. More women less than 30 were infected with HR-HPV and multiple genotypes regardless of the L-Pap reading. AMP-HPV and LA-HPV testing had an overall raw agreement with each other of 84.2% [Kappa 0.66] and each had agreement of 94% with HC2 testing of 133 samples [Kappa 0.86/0.87]. AMP-HPV agreement between two laboratories was better at 93% [Kappa 0.84] compared to 76.1% [Kappa 0.40] when extraction was standardized. Conclusion It is feasible to perform AMP-HPV and LA-HPV on SurePath samples to detect HR-HPV genotypes. HC2, AMP-HPV and LA-HPV showed strong agreement. The extraction component of the AMP-HPV assay needs careful attention to yield consistent results.