Infection of Fetal Membranes with Mycobacterium tuberculosis and Tissue Processing to Isolate RNA for Expression
Samantha Yong-Mendoza1,2, Mónica Selena Fonseca-Pérez1,2, Oscar Villavicencio-Carrisoza1
1Department of Immunobiochemistry, Instituto Nacional de Perinatologia Isidro Espinosa de los Reyes, Mexico City, Mexico.
Abstract:
This chapter outlines the methodology employed to infect the chorionic and amniotic membranes with Mycobacterium tuberculosis during pregnancy. Particularly, congenital tuberculosis, a rare and serious condition associated with cases in neonates and reactivation of latent tuberculosis in pregnant mothers, is interesting to study. Understanding the mechanisms of infection and the response of fetal membranes is crucial for developing effective treatments in these cases, which will promote better neonatal and maternal health in situations of tuberculosis during pregnancy. Establishing a standardized infection model in the chorioamniotic membranes is imperative, followed by a treatment protocol for isolating both cellular and mycobacterial RNA. This will enable the expression analysis during the maternal-fetal interface interaction with M. tuberculosis. The proposed methodology might be invaluable for qRT-PCR, microarrays, and sequencing research.
Insights
This study details a method to infect pregnancy membranes with Mycobacterium tuberculosis, aiding research into congenital tuberculosis. This infection model is key for understanding maternal-fetal interactions and improving neonatal and maternal health outcomes.
Area of Science:
- Obstetrics and Gynecology
- Infectious Diseases
- Maternal-Fetal Medicine
Background:
- Congenital tuberculosis is a rare but severe condition in neonates, often linked to latent tuberculosis reactivation in pregnant individuals.
- Understanding Mycobacterium tuberculosis infection mechanisms within fetal membranes is critical for effective treatment strategies.
- Improved maternal and neonatal health outcomes are a priority in managing tuberculosis during pregnancy.
Purpose of the Study:
- To outline a methodology for infecting chorionic and amniotic membranes with Mycobacterium tuberculosis during pregnancy.
- To establish a standardized infection model for studying maternal-fetal interface interactions with M. tuberculosis.
- To facilitate research into congenital tuberculosis and its impact on pregnancy.
Main Methods:
- Infection of chorionic and amniotic membranes with Mycobacterium tuberculosis in a pregnancy model.
- Development of a treatment protocol for isolating cellular and mycobacterial RNA.
- Preparation for expression analysis using techniques like qRT-PCR, microarrays, and sequencing.
Main Results:
- A standardized methodology for M. tuberculosis infection of fetal membranes has been established.
- The protocol allows for the isolation of both host and pathogen RNA for downstream analysis.
- The model is suitable for investigating gene expression at the maternal-fetal interface during tuberculosis infection.
Conclusions:
- The proposed methodology provides a valuable tool for studying congenital tuberculosis.
- This research is crucial for advancing our understanding of maternal-fetal interactions during M. tuberculosis infection.
- The model supports the development of targeted treatments to improve maternal and neonatal health outcomes.
