2014, Número 3
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Rev Cubana Med Trop 2014; 66 (3)
Normalización de un método inmunoquímico para detectar Papilomavirus humano tipo 16 en lesiones cérvico-uterinas
Torres LG, Soto BY, Capó PV, López FLX, de la Torre JAI, Goicolea MA, Limonta VD, Pérez RA, Govín CA, Morier DL, Caballero LY, Rodríguez SH, Álvarez GRI, Calvo PA, Escobar PX, Limia LCM, Kourí CV
Idioma: Español
Referencias bibliográficas: 27
Paginas: 433-446
Archivo PDF: 271.49 Kb.
RESUMEN
Introduction: Human Papillomavirus (HPV) infection is the necessary condition for
the occurernce and development of cervical cancer. The high oncogenic risk
genotypes are the responsible for this type of neoplasia and the most frequent is
HPV 16 that affects roughly 60 % of cases. Commercial kits for HPV detection are
expensive for resource-poor countries, which suggests the search for alternative
throguh non-expensive simple protocoles.
Objectives: to standardize an immunochemical method for the detection of HPV 16
L1 antigen in cervical samples of patients with squamous intraepithelial lesions and
to determine the diagnostic coincidence between the immunochemical method and
the real-time polymerase chain reaction to estimate the usefulness of this method
for the detection of cervical infection with this viral genotype.
Methods: three immunostaining methods (Two-Step Indirect Immunoperoxidase,
Labelled Streptavidin-Biotin and Enhanced Polymer) were compared in terms of
analytical sensitivity, nonspecific background staining and time of completion, for
the detection of protein L1 of HPV-16 in a cell line derived from human cervical
carcinoma and clinical samples from uterine cervix. The optimized protocol was
applied to 82 cervical samples from women aged 30-59 years with squamous
intraepithelial lesions and to 10 samples of sexually active women without previous
signals of positive cytology. The presence of type 16 HPV was also detected with
the aid of RT-PCR.
Results: the Streptavidin-Biotin system was the most sensitive and specific. The
diagnostic agreement between the immunochemical method and the real-time
polymerase chain reaction reached 98.6 %, sensitivity was 98.57 % and specificity
was 91.67 %, with positive and negative predictive values above 90 %.
Conclusions: the validity of the immunochemical method as a confirmatory test for
infection by HPV-16 has been demonstrated. The normalized immunochemical method proved to be a sensitive, simple, relatively fast method to detect HPV from clinical samples of cervical cells. Furthermore, this method provides information quickly, avoiding the use of invasive methods in patients.
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