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Bacterial-based systems for expression and purification of recombinant Lassa virus proteins of immunological relevance
Authors:Luis M Branco  Alex Matschiner  Joseph N Fair  Augustine Goba  Darryl B Sampey  Philip J Ferro  Kathleen A Cashman  Randal J Schoepp  Robert B Tesh  Daniel G Bausch  Robert F Garry  Mary C Guttieri
Institution:1. Diagnosis – Molecular Laboratory, University of Caxias do Sul, Caxias do Sul, Rio Grande do Sul, Brazil
3. Biotechnology Institute, University of Caxias do Sul, Caxias do Sul, Rio Grande do Sul, Brazil
4. Outpatient Clinic of Genital Pathology, Department of Clinical Medicine, University of Caxias do Sul, Caxias do Sul, Rio Grande do Sul, Brazil
2. Pathology Medical Laboratory, Department of Health and Biomedical Science, University of Caxias do Sul, Caxias do Sul, Rio Grande do Sul, Brazil
Abstract:This paper aimed at studying the transplacental transmission of HPV and looking at the epidemiological factors involved in maternal viral infection. The following sampling methods were used: (1) in the pregnant woman, (a) genital; (b) peripheral blood; (2) in the newborn, (a) oral cavity, axillary and inguinal regions; (b) nasopharyngeal aspirate, and (c) cord blood; (3) in the placenta. The HPV DNA was identified using two methods: multiplex PCR of human β-globin and of HPV using the PGMY09 and PGMY11 primers; and nested-PCR, which combines degenerated primers of the E6/E7 regions of the HPV virus, that allowed the identification of genotypes 6/11, 16, 18, 31, 33, 42, 52 and 58. Transplacental transmission was considered when type-specific HPV concordance was found between the mother, the placenta and the newborn or the mother and cord blood. The study included 49 HPV DNA-positive pregnant women at delivery. Twelve placentas (24.5%, n = 12/49) had a positive result for HPV DNA. Eleven newborn were HPV DNA positive in samples from the nasopharyngeal or buccal and body or cord blood. In 5 cases (10.2%, n = 5/49) there was HPV type-specific agreement between genital/placenta/newborn samples. In one case (2%, n = 1/49) there was type specific HPV concordance between genital/cord blood and also suggested transplacental transmission. A positive and significant correlation was observed between transplacental transmission of HPV infection and the maternal variables of immunodepression history (HIV, p = 0.011). In conclusion the study suggests placental infection in 23.3% of the cases studied and transplacental transmission in 12.2%. It is suggested that in future HPV DNA be researched in the normal endometrium of women of reproductive age. The possible consequence of fetal exposure to HPV should be observed.
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