A review of past, present and emerging techniques for detection of Mycoplasma pneumoniae.

Mycoplasma pneumoniae is a significant pathogen causing respiratory infections globally, with diagnosis complicated by the organism's unique biology and nonspecific clinical presentation. Its distinct biology, including the absence of a cell wall and ambiguous clinical presentation, challenges early and accurate diagnosis, which is critical for guiding optimal antimicrobial therapy and limiting antibiotic resistance. This study summarizes the progression of diagnostic procedures from traditional culture and serology to modern molecular techniques such as real-time PCR, multiplex PCR panels, isothermal amplification methods like SAT, LAMP, and emerging techniques such as NGS and CRISPR. The paper underlines the advantages of molecular approaches over culture and serology in terms of sensitivity, specificity, and turnaround time, focusing on FDA/CE-cleared multiplex PCR systems and quick antigen detection kits with silver amplification as valuable clinical tools. Emerging CRISPR-based diagnostics offer a wonderful opportunity for quick, ultrasensitive, and point-of-care diagnosis. Despite the variety of available assays, issues such as detecting asymptomatic carriage, distinguishing active infection, and ensuring accessibility in low-resource settings persist. The increased global incidence of macrolide-resistant M. pneumoniae strains, as well as the overlap of respiratory symptoms with other diseases, highlight the need for specific, quick, and reliable diagnostic tests. This review advocates for the incorporation of innovative molecular diagnostics into standard clinical workflows to improve early identification, optimize treatment methods, and improve patient outcomes.