Development and Validation of a Quantitative Synthetic Molecular Standard for Coxiella burnetii

Q fever is a highly infectious zoonotic disease caused by Coxiella burnetii, a group B biological warfare agent found worldwide that infects humans and a wide range of domestic and wild animals. This disease is associated with reproductive disorders in animals; in humans, symptoms can be mild or severe and may progress to pneumonia or chronic complications such as endocarditis, meningoencephalitis, and osteomyelitis. To control zoonotic transmission, accurate and sensitive detection is critical. Currently, real-time PCR assays are routinely used to ensure rapid detection and quantification; however, the accuracy and reproducibility of these assays are limited by the lack of precisely quantified controls. While genomic DNA can be used as a standard for these assays, C. burnetii is difficult to cultivate as it is a slow growing obligate intracellular pathogen that requires BSL-3 facilities. To address this need, ATCC developed a quantitative synthetic molecular standard for C. burnetti (ATCC BAA-4000SD). As a proof-of-concept, we tested the functionality of the standard via qPCR with a published primer and probe set, and protocol. To make high-containment pathogens more accessible, this approach was also extended to Nipah virus and Lassa virus.