Highly Sensitive Detection and Genotyping of HPV by PCR Multiplex and Luminex Technology in a Cohort of Colombian Women with Abnormal Cytology
Dabeiba A García1, Angel Cid-Arregui*, 2, Markus Schmitt3, Marcos Castillo4, 5, Ignacio Briceño4, Fabio A. Aristizábal66
1 Centro de Investigaciones Odontológicas, Pontificia Universidad Javeriana, Bogota, Colombia
2 Translational Immunology Unit, German Cancer Research Center (DKFZ), Heidelberg, Germany
3 Division of Genome Modifications and Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg, Germany
4 Facultad de Medicina, Universidad de La Sabana, Colombia
5 Dinamica-IPS, Bogotá, Facultad de Ciencias, Universidad Nacional de Colombia, Colombia
6 Departamento de Farmacia, Facultad de Ciencias, Universidad Nacional de Colombia, Colombia
Cancer of the uterine cervix (CC) is the second most common cancer in women worldwide. In Colombia, CC is the second most frequent cancer among the entire women population and the first among women aged between 15 and 44 years, with an estimated incidence of 24.9 cases/100,000 inhabitants. The main risk factor is infection with one or more high-risk human papillomavirus (HPV) types. The aim of this study was to estimate the genotype-specific prevalence of human papillomavirus (HPV) DNA in patients with cervical pathology using the multiplex PCR and Luminex xMAP technology. In addition, we compared genotyping with Luminex xMAP and with Reverse Line Blot (RLB). A cohort of 160 patients participated in the study, of which 25.6% had no cervical lesions, 35% presented cervical intraepithelial neoplasia of grade I (CIN I), 10% CIN II, 20.6% CIN III and 8.8% CC. The most frequent viral types in all lesion grades were HPV16 and HPV18. Infections by a unique virus were less frequent (19.4%) than multiple infections (80.6%). Single infections were found in 22% of women with no cervical lesions, and in 14.3% of CIN I, 18.7% CIN II, 21.2% CIN III and 28.6% of CC. Multiple infections were observed in 78.0% of cervical samples with negative histopathologic diagnosis, and in 85.7% of CIN I, 81.2% CIN II, 78.8% CIN III and 71.4% CC. All samples analyzed with Luminex xMAP were HPV-positive, while we could detect HPV in only 48.8% of cases with RLB. Of the samples positive by both methods, there was a 67.2% correlation in the viral type(s) detected. In conclusion, Luminex suspension array showed a remarkably higher sensitivity compared with RLB. Multiple infections were unexpectedly common, being HPV types 16 and 18 the most prevalent in all histopathologic grades.
Keywords: Human papillomavirus (HPV), CIN, cervical cancer, reverse line blot (RLB), luminex xMAP, pap smear, abnormal cytology..
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* Address correspondence to this author at the Translational Immunology Group D015, German Cancer Research Center (DKFZ), Im Neuenheimer Feld 280, D-69120 Heidelberg, Germany; Tel: +49 6221 567109; Fax: +49 6221 564776; E-mail: firstname.lastname@example.org