Amplification of early genes of Human Papilloma Virus targeting nine virus genotypes. Mérida, Venezuela

Abstract

Background: In 2007, Gaynor et al. discovered a previously unidentified human polyomavirus in respiratory secretions obtained from human patients with symptoms of acute respiratory tract infection. T his new virus was designated WU polyomavirus.To investigate the frequency of it infections in GuangDong, China, We sought to describe the detection and clinical characterization of WU Polyomavirus with infection in Children.Methods: From July 2008 through June 2009, nasopharyngeal aspirates were collected from 771 child ren who were hospitalized with acute lower respiratory tract infection in Second Affiliated Hos pital of Shantou University Medical College, and 82 were asymptomatic who visited the wellb eing clinic .WU Polyomaviruses was detected by using PCR technology and was identified by using DNA sequences. All WU polyomavirus positive specimens were screened for 9 comme n viruses (influenza A and B; RSV; PIV 1 and 3; human metapneumovirus; human bocavirus; adenovirus; rhinovirus)by using PCR or RT-PCR.Results: In this study, fifteen of the 771 tested specimens with acute lower respiratory tract infection were positive fo r WU polyomavirus, the positive rate was 1.95%. and all of the asymptomatic children who vis ited the wellbeing clinic were negative. Positive specimens were noted for patients 2 months to 48 months of age, the median age was 18.8 months. Of the 15 WU Polyomaviruses positive patients, 9 (60%) were male.,6(40%) were female.WU polyomavirus was the sole virus detected in 9 spe cimens(60%) from patients with acute respiratory tract infection. WU polyomavirus were asso ciated with the coinfection of another respiratory virus in 6 of 15 (40%), most frequently with RSV (n = 4),follo wed by adenovirus (n = 1) and rhinovirus (n = 1). The most common clinical findings in the pa tients with WU polyomavirus are cough,fever, wheezing.The most frequent diagnoses were pneumonia (n = 8),bronchiolitis(n = 4), upper respiratory tract infections (n = 2) and bronchitis(n = 1).A severe case was complication with viral encephalitis.Conclusion: We suggests that WU Polyomaviruses may be a respiratory pathogen because WU polyomav irus was the sole virus detected in 9 specimens from patients with respiratory illness and all of the asymptomatic were negative.The most common clinical findings are cough and wheezing .Young children are the main target. And the pathogenetic condition are generally mild. More comprehensive studies are required to prove these viruses are agents causing respiratory disease.

Evaluación de la expresión de ARNm de genes virales E2, E6 y E7 como marcadores predictivos de progresión en lesiones producidas por VPH 16

“…El proceso de genotipificación consistió en una PCR multiplex anidada en dos pasos; en el primer paso se realizó una ampliación utilizando cebadores de consenso GP5+/ GP6+ y MY09/MY11 que reconocen secuencias comunes del Gen L1 en todos los VPH, Investigación Clínica 59(4): 2018 esto permite la detección del genoma viral en la muestra (15,16). Posteriormente se realizó la tipificación, consistente en una PCR multiplex anidada de 2 rondas programadas en un termociclador convencional TC-3000X.…”

“…En la primera ronda se amplificó la región E6/E7, facilitando y mejorando los resultados en la segunda ronda, en la cual se utilizaron varios conjuntos de cebadores permitiendo la tipificación de 19 genotipos. Esta metodología no solo logra determinar el genotipo de VPH presente, también permite detectar infecciones por múltiples genotipos de VPH (coinfección) (15)(16)(17).…”

“…La amplificación de la secuencia específica del gen E7 del VPH 16 se realizó por medio de la PCR convencional utilizando la pareja de cebadores y la programación estandarizados para el termociclador TC-3000X en (1,16,17). En la amplificación de la secuencia específica del gen E6 del VPH 16, la pareja de cebadores utilizados y la programación del termociclador TC-3000X se realizó en base a trabajos consultados (15)(16)(17).…”

“…En la amplificación de la sección específica del gen E2 del VPH 16 la pareja de cebadores y la programación del termociclador TC-3000X fueron los mismos que el trabajo consultado de (16).…”