Molecular biology Archives - Revista Brasileira de Ginecologia e Obstetrícia
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2015;37(5):203-207
DOI 10.1590/SO100-720320150005293
To determine the frequency of Human Papillomavirus (HPV) in the placenta, in the
colostrum and in the umbilical cord blood of parturient women and their newborns
assisted at the Clinic of Gynecology and Obstetrics of the University Hospital of
Rio Grande (RS), Brazil.
Biopsies were collected from 150 placentas on the maternal side, 150 on the fetal
side, 138 samples of umbilical cord blood and 118 of the colostrum. The placenta
biopsies were collected from the central and peripheral portions. DNA was
extracted according to the manufacturer's protocol and to a reference found in the
literature. HPV was detected by the nested polymerase chain reaction (PCR-Nested)
using primers MY09/11 and GP5/GP6. Genotyping was performed by direct sequencing.
The participants responded to a self-applied questionnaire with demographic and
clinical data, in order to characterize the sample.
HPV was detected in 4% (6/150) of cases on the mother's side of the placentas, in
3.3% (5/150) on the fetal side, in 2.2% (3/138) in umbilical cord blood and in
0.84% (1/118) in colostrum samples. The vertical transmission rate was 50%. HPV-6
was the low-risk genotype found (60%) and the high-risk genotypes were HPV-16 and
HPV-18 (20% each).
These results suggest that HPV can infect the placenta, the colostrum and the
umbilical cord blood.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2015;37(5):203-207
DOI 10.1590/SO100-720320150005293
To determine the frequency of Human Papillomavirus (HPV) in the placenta, in the
colostrum and in the umbilical cord blood of parturient women and their newborns
assisted at the Clinic of Gynecology and Obstetrics of the University Hospital of
Rio Grande (RS), Brazil.
Biopsies were collected from 150 placentas on the maternal side, 150 on the fetal
side, 138 samples of umbilical cord blood and 118 of the colostrum. The placenta
biopsies were collected from the central and peripheral portions. DNA was
extracted according to the manufacturer's protocol and to a reference found in the
literature. HPV was detected by the nested polymerase chain reaction (PCR-Nested)
using primers MY09/11 and GP5/GP6. Genotyping was performed by direct sequencing.
The participants responded to a self-applied questionnaire with demographic and
clinical data, in order to characterize the sample.
HPV was detected in 4% (6/150) of cases on the mother's side of the placentas, in
3.3% (5/150) on the fetal side, in 2.2% (3/138) in umbilical cord blood and in
0.84% (1/118) in colostrum samples. The vertical transmission rate was 50%. HPV-6
was the low-risk genotype found (60%) and the high-risk genotypes were HPV-16 and
HPV-18 (20% each).
These results suggest that HPV can infect the placenta, the colostrum and the
umbilical cord blood.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2011;33(10):315-320
DOI 10.1590/S0100-72032011001000008
PURPOSE: to compare three methods for the detection of HPV infection and to determine the prevalence of the genotypes found. METHODS: a total of 120 cervical scrape samples from patients with cervical intraepithelial neoplasia were analyzed by the conventional polymerase chain reaction using the MY09/11 and GP05+/06+ primers, and by the Nested polymerase chain reaction. The samples were subjected to DNA amplification with the GH20 and PC04 primers (β-globin) to verify DNA quality and also by polymerase chain reaction and Nested polymerase chain reaction. The amplicons were visualized in 1.2% agarose gel stained with Blue Green Loading Dye I. Positive samples also were sequenced using the automatic DNA sequencer "MegaBACE 1000". The Χ2 and Fisher tests were used for statistical analysis with the level of significance set at 5%. RESULTS: fifteen samples were eliminated from the study because they failed to amplify the β-globin gene. Of the remaining samples, 40% (42/105) were positive using primers MY09/11, 98% (103/105) using primers GP05+/06+, and 92% (97/105) using Nested-PCR. With the MY09/11 and GP05+/06+ techniques, it was possible to obtain 100% HPV-positive samples. In this study, the prevalence of the genotypes found was 57, 23, 5, 4 and 3% for HPV genotypes 16, 18, 31, 33 and 56, respectively. HPVs 67 and 83 were present in 2%, and genotypes 6, 11, 58 and candHPV85 were present in 1% each. The prevalence of the more common genotypes (HPV 16 and 18) in this study agrees with that reported worldwide (IC95%=0.4657-0.8976). CONCLUSIONS: to obtain more reliable results, it is necessary the use of more than one primer system to detect HPV infections. We believe that the three techniques studied are important and suitable for the clinical diagnosis of HPV, when they are appropriately combined.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2011;33(10):315-320
DOI 10.1590/S0100-72032011001000008
PURPOSE: to compare three methods for the detection of HPV infection and to determine the prevalence of the genotypes found. METHODS: a total of 120 cervical scrape samples from patients with cervical intraepithelial neoplasia were analyzed by the conventional polymerase chain reaction using the MY09/11 and GP05+/06+ primers, and by the Nested polymerase chain reaction. The samples were subjected to DNA amplification with the GH20 and PC04 primers (β-globin) to verify DNA quality and also by polymerase chain reaction and Nested polymerase chain reaction. The amplicons were visualized in 1.2% agarose gel stained with Blue Green Loading Dye I. Positive samples also were sequenced using the automatic DNA sequencer "MegaBACE 1000". The Χ2 and Fisher tests were used for statistical analysis with the level of significance set at 5%. RESULTS: fifteen samples were eliminated from the study because they failed to amplify the β-globin gene. Of the remaining samples, 40% (42/105) were positive using primers MY09/11, 98% (103/105) using primers GP05+/06+, and 92% (97/105) using Nested-PCR. With the MY09/11 and GP05+/06+ techniques, it was possible to obtain 100% HPV-positive samples. In this study, the prevalence of the genotypes found was 57, 23, 5, 4 and 3% for HPV genotypes 16, 18, 31, 33 and 56, respectively. HPVs 67 and 83 were present in 2%, and genotypes 6, 11, 58 and candHPV85 were present in 1% each. The prevalence of the more common genotypes (HPV 16 and 18) in this study agrees with that reported worldwide (IC95%=0.4657-0.8976). CONCLUSIONS: to obtain more reliable results, it is necessary the use of more than one primer system to detect HPV infections. We believe that the three techniques studied are important and suitable for the clinical diagnosis of HPV, when they are appropriately combined.