Last data update: Jun 24, 2024. (Total: 47078 publications since 2009)
Records 1-1 (of 1 Records) |
Query Trace: Linscott KB [original query] |
---|
sodC-based real-time PCR for detection of Neisseria meningitidis.
Dolan Thomas J , Hatcher CP , Satterfield DA , Theodore MJ , Bach MC , Linscott KB , Zhao X , Wang X , Mair R , Schmink S , Arnold KE , Stephens DS , Harrison LH , Hollick RA , Andrade AL , Lamaro-Cardoso J , de Lemos AP , Gritzfeld J , Gordon S , Soysal A , Bakir M , Sharma D , Jain S , Satola SW , Messonnier NE , Mayer LW . PLoS One 2011 6 (5) e19361 ![]() Real-time PCR (rt-PCR) is a widely used molecular method for detection of Neisseria meningitidis (Nm). Several rt-PCR assays for Nm target the capsule transport gene, ctrA. However, over 16% of meningococcal carriage isolates lack ctrA, rendering this target gene ineffective at identification of this sub-population of meningococcal isolates. The Cu-Zn superoxide dismutase gene, sodC, is found in Nm but not in other Neisseria species. To better identify Nm, regardless of capsule genotype or expression status, a sodC-based TaqMan rt-PCR assay was developed and validated. Standard curves revealed an average lower limit of detection of 73 genomes per reaction at cycle threshold (C(t)) value of 35, with 100% average reaction efficiency and an average R(2) of 0.9925. 99.7% (624/626) of Nm isolates tested were sodC-positive, with a range of average C(t) values from 13.0 to 29.5. The mean sodC C(t) value of these Nm isolates was 17.6+/-2.2 (+/-SD). Of the 626 Nm tested, 178 were nongroupable (NG) ctrA-negative Nm isolates, and 98.9% (176/178) of these were detected by sodC rt-PCR. The assay was 100% specific, with all 244 non-Nm isolates testing negative. Of 157 clinical specimens tested, sodC detected 25/157 Nm or 4 additional specimens compared to ctrA and 24 more than culture. Among 582 carriage specimens, sodC detected Nm in 1 more than ctrA and in 4 more than culture. This sodC rt-PCR assay is a highly sensitive and specific method for detection of Nm, especially in carriage studies where many meningococcal isolates lack capsule genes. |
- Page last reviewed:Feb 1, 2024
- Page last updated:Jun 24, 2024
- Content source:
- Powered by CDC PHGKB Infrastructure