Hello, my name is Lavapoku Afrifah, and welcome to my OMED 2024 virtual poster presentation. I am a fourth year at Burrell College of Osteopathic Medicine, and I want to thank you again for joining me for my presentation on Lamping Up Urogenital Diagnostics, a multiplex assay for simultaneous Mycoplasma Genitalium and Group B Streptococcus detection. So to begin, Mycoplasma Genitalium, or MGen, and Group B Streptococcus, or GBS, pose significant public health challenges. MGen is a sexually transmitted infection linked to non-gonococcal urethritis in men and premature rupture of membranes in women during pregnancy. GBS, while not exclusively sexually transmitted, can cause severe infection in newborns, pregnant women, and immunocompromised individuals. Accurate and timely diagnosis of both pathogens are crucial for effective treatment and prevention of complications. Traditional diagnostic methods for MGen and GBS often include complex laboratory procedures such as culturing and polymerase chain reaction, or PCR. These methods can be time-consuming, expensive, and require specialized equipment, limiting their availability in resource-constrained settings. Furthermore, the fastidious nature of MGen can make its culture quite challenging, often leading to delayed or inaccurate diagnosis. So we designed a multiplex loop-mediated isothermal amplification, or LAMP, assay targeting the PDHD gene of Mycoplasma and the CYP gene of Group B Streptococcus. LAMP is a rapid and isothermal nucleic acid amplification technique that offers several advantages over conventional PCR, including its ability to be performed at a constant temperature without the need for thermocycling. We utilized primer design software from New England Biolabs to create highly specific LAMP primers for both pathogens. The LAMP assay was initially optimized for colorimetric detection and subsequently adapted for lateral flow assay, or LFA, visualization. Our novel LAMP assay demonstrated high sensitivity and specificity for the detection of MGen and GBS individually and in multiplex. We successfully optimized the LAMP reaction conditions, identifying an optimal temperature of 66.7 degrees Celsius for the most robust amplification of both target organisms. The PDHD color LAMP achieved a limit of detection of 15.97 genomic copies of Mycoplasma genitalium within 30 minutes and a limit of detection for Strep agalactiae of 427 genomic copies. The LFA platform enabled rapid and visual detection of both MGen and GBS, making it suitable for point-of-care applications. In conclusion, the development of this multiplex LAMP assay represents a significant step towards improved diagnosis and management of Mycoplasma genitalium and Group B Streptococcus infections. By enabling rapid and accurate detection at the point-of-care, this assay has the potential to enhance patient care and public health outcomes. Further studies will focus on evaluating the clinical performance of this assay in a broader patient population. Thank you again for joining me for my presentation on LAMPing Up Urogenital Diagnostics, a Multiplex Assay for Simultaneous MGen and Group B Streptococcus Detection. My references can be found in the QR code at the bottom of the screen. Thank you again and have a great rest of your day.

LAMP assay

Mycoplasma Genitalium

Group B Streptococcus

diagnostic accessibility

point-of-care detection