Last data update: Aug 15, 2025. (Total: 49733 publications since 2009)
| Records 1-1 (of 1 Records) |
| Query Trace: DeLaney AA [original query] |
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| Isothermal detection of Mycoplasma pneumoniae directly from respiratory clinical specimens
Petrone BL , Wolff BJ , DeLaney AA , Diaz MH , Winchell JM . J Clin Microbiol 2015 53 (9) 2970-6 Mycoplasma pneumoniae is a leading cause of community-acquired pneumonia (CAP) across patient populations of all ages. We have developed a loop-mediated isothermal amplification (LAMP) assay that enables rapid, low-cost detection of M. pneumoniae from nucleic acid extracts and directly from various respiratory specimen types. The assay implements calcein to facilitate simple visual readout of positive results in approximately one hour, making it ideal for use in primary care facilities and resource-poor settings. The analytical sensitivity of the assay was determined to be 100 fg by testing serial dilutions of target DNA ranging from 1 ng to 1 fg per reaction, and no cross-reactivity was observed against 17 other Mycoplasma species, 27 common respiratory agents, or human DNA. We demonstrate the utility of this assay by testing nucleic acid extracts (n=252) and un-extracted respiratory specimens (n=72) collected during M. pneumoniae outbreaks and sporadic cases occurring in the United States from February 2010 to January 2014. The sensitivity of the LAMP assay was 88.5% when tested on extracted nucleic acid and 82.1% when evaluated on un-extracted clinical specimens, compared to a validated real-time PCR test. Further optimization and improvements to this method may lead to a rapid, cost-efficient laboratory test for M. pneumoniae detection that is more widely available to primary care facilities, ultimately facilitating prompt detection and appropriate response to potential M. pneumoniae outbreaks and clusters within the community. |
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