Article Impact Level: HIGH Data Quality: STRONG Summary of Journal of Clinical Investigation. https://doi.org/10.1172/JCI177241 Dr. Brady M. Youngquist et al.
Points
- The study developed an ultrasensitive RT-PCR-coupled CRISPR assay to improve the detection of Pneumocystis jirovecii pneumonia (PCP) using noninvasive samples like oropharyngeal swabs and serum.
- Current PCP diagnosis relies on bronchoalveolar lavage (BAL), which is invasive and difficult to obtain, especially in infants, prompting the need for alternative diagnostic approaches.
- In human studies, the CRISPR assay showed 96.3% sensitivity in infant swabs (66.7% for RT-qPCR) and 93.3% sensitivity in adult serum samples (26.7% for RT-qPCR), with 100% specificity.
- The assay successfully detected Pneumocystis RNA in serum samples throughout the infection period in a mouse model, demonstrating its potential for early and reliable diagnosis.
- This high-accuracy, non-invasive method could reduce dependence on BAL, making PCP diagnosis more accessible and efficient, particularly for pediatric and immunocompromised patients.
Summary
This study aimed to enhance the diagnostic workflow for Pneumocystis jirovecii pneumonia (PCP) by developing an ultrasensitive RT-PCR-coupled CRISPR assay for improved detection using non-invasive samples like oropharyngeal swabs and serum. Traditional diagnosis of PCP relies heavily on bronchoalveolar lavage (BAL), which is difficult to obtain, particularly in infants. The authors hypothesized that CRISPR could improve the sensitivity of assays, allowing for more accessible and efficient diagnostics. The study employed mouse models and human specimens to validate the assay’s efficacy in detecting Pneumocystis RNA in various sample types.
In the mouse model, the RT-PCR CRISPR assay detected Pneumocystis RNA in serum samples throughout the infection period. In human studies, the CRISPR assay performed significantly better than traditional RT-qPCR. Specifically, oropharyngeal swabs from infants with PCP demonstrated a sensitivity of 96.3% (compared to 66.7% with RT-qPCR) and specificity of 100% (compared to 90.6% with RT-qPCR). Additionally, the CRISPR assay achieved higher sensitivity in adult serum samples, detecting P. jirovecii with 93.3% sensitivity, while RT-qPCR only achieved 26.7%. These results highlight the CRISPR assay’s potential to identify active P. jirovecii infections more reliably than current methods.
The findings suggest that this CRISPR-enhanced RT-PCR assay could transform PCP diagnosis, particularly in pediatric patients where swabs are routinely collected, and serum is easier to obtain than BAL. With its high sensitivity and specificity for active infection, this approach could reduce the reliance on invasive procedures like BAL, thereby improving diagnostic accuracy and speed in pediatric and adult populations.
Link to the article: https://www.jci.org/articles/view/177241
References Youngquist, B. M., Mnguni, A. T., Pungan, D., Lai, R. P. J., Dai, G., Ng, C. F., Samson, A., Abdelgaliel, Y., Lyon, C. J., Ning, B., Husain, S., Wasserman, S., Kolls, J. K., & Hu, T. Y. (2025). CRISPR-mediated detection of Pneumocystis transcripts in bronchoalveolar, oropharyngeal, and serum specimens for Pneumocystis pneumonia diagnosis. Journal of Clinical Investigation. https://doi.org/10.1172/JCI177241
