RESUMO
The Hepatitis B surface antigen (HBsAg) is the hallmark of HBV infection. Detection of antibodies to HBs and the core (ie, HBsAg and HBcAb) are primary serological algorithms in the laboratory diagnosis of HBV. Detection of HBsAg DNA is an important supplement to serological diagnosis especially in clinical cases. Simultaneous amplification of internal cellular controls is a good indicator of sample quality. Human ß-globin is a well characterized housekeeping gene (HKG) that is often applied as internal controls (IC) in molecular diagnosis. In this study, individual plasmid clones of the human ß-globin and HBs genes were constructed. These plasmid constructs have been applied to characterize a multiplex PCR assays for HBs and ß-globin genes. The findings suggest detection limits of less than 10 genome copies of either template In vitro using conventional and multiplex PCR conditions. Under the multiplex conditions, co-amplification of ß-globin and HBsAg DNA had a resultant effect on assay sensitivity. This study further highlights the importance of molecular diagnosis in HBV infectious individuals. If fully optimized, this assay could provide a possible diagnostic complement to serological detection in developing countries.
Assuntos
Antígenos de Superfície da Hepatite B/genética , Vírus da Hepatite B/isolamento & purificação , Hepatite B/diagnóstico , Reação em Cadeia da Polimerase Multiplex/métodos , Globinas beta/genética , Células A549 , Linhagem Celular Tumoral , DNA Viral/genética , Genes Essenciais , Células HEK293 , Hepatite B/virologia , Vírus da Hepatite B/genética , Humanos , Técnicas de Diagnóstico Molecular/métodos , Plasmídeos/genética , Sensibilidade e EspecificidadeRESUMO
Regional variations exist in HPV prevalence worldwide despite reports of high prevalence rates among African women. Limited data on genital HPV prevalence necessitated this study with the aim of determining the prevalence of genital HPV and associated risk factors among women in Lagos, Nigeria. Exfoliated cervical cells were collected with consent from 165 women using a cervical brush. Viral DNA was extracted and amplified by nested PCR using two sets of consensus primers (MY09/11 and GP5+/6+). An unconditional logistic regression model was used to identify predictors of HPV positivity. The HPV prevalence was 81.82% in all women and 87.59% in women with normal cytology. The risk of HPV infection was significantly increased among women who had a history of STI (odds ratio (OR) 3.94; 95% confidence interval (CI): 1.51-10.25, p = 0.005) while there was a significantly reduced risk of HPV infection among those who used condoms (odds ratio (OR) 3.94; 95% confidence interval (CI): 0.18-0.91, p = 0.03). The HPV prevalence observed shows an increased transmission of the virus in Lagos, Nigeria. Therefore, there is a need for intense public awareness and the implementation of early detection tests, treatment, and vaccination to prevent an increase in cervical cancer cases in Lagos, Nigeria.
RESUMO
Investment in Africa over the past year with regards to SARS-CoV-2 genotyping has led to a massive increase in the number of sequences, exceeding 100,000 genomes generated to track the pandemic on the continent. Our results show an increase in the number of African countries able to sequence within their own borders, coupled with a decrease in sequencing turnaround time. Findings from this genomic surveillance underscores the heterogeneous nature of the pandemic but we observe repeated dissemination of SARS-CoV-2 variants within the continent. Sustained investment for genomic surveillance in Africa is needed as the virus continues to evolve, particularly in the low vaccination landscape. These investments are very crucial for preparedness and response for future pathogen outbreaks. One-Sentence SummaryExpanding Africa SARS-CoV-2 sequencing capacity in a fast evolving pandemic.