Evaluation of a Real-Time PCR Assay to Detect Coxiella burnetii

2006 ◽  
Vol 1078 (1) ◽  
pp. 563-565 ◽  
Author(s):  
S. R KLEE ◽  
H. ELLERBROK ◽  
J. TYCZKA ◽  
T. FRANZ ◽  
B. APPEL
2017 ◽  
Vol 4 (2) ◽  
pp. 11
Author(s):  
Mojtaba Bonyadian ◽  
Hamdollah Moshtaghi ◽  
Hamidreza Kazemeini

Coxiella burnetii is the causative agent of Q-fever, a widespread zoonosis. In domestic animals infection remains either asymptomatic or presents as infertility or abortion. Clinical presentation in humans can range from mild flu-like illness to acute pneumonia and hepatitis. In humans serology is the gold standard for diagnosis but is inadequate for early case detection, so real-time PCR and nested-PCR assays were developed in this study to measure amounts of C.burnetii shed in milk. Our study was to assess the sensitivity of the realtime PCR and nested-PCR for detection of Coxiella burnetii in bovine bulk milk samples from dairy herds in 3 provinces (Chaharmahal and Bakhtiari , Isfahan and Yazd) of Iran. In the present study, 300 bulk milk samples from 89 dairy cattle herds were tested for C. burnetii using real-time PCR and nested-PCR assays. The animals which their milk samples collected for this study were clinically healthy. In total, 74 of 300 (24.7%) cow milk samples were positive in real-time PCR assay and 26 of 300 (8.7%) samples were positive in nested-PCR assay. McNemar test shows a significant difference in detection of C. burnetii between real-time PCR and nested-PCR. Also the results of this study indicate those clinically healthy dairy cows are important sources of C. burnetii infection in Iran.


2016 ◽  
Vol 55 (2) ◽  
pp. 180-184 ◽  
Author(s):  
Solène Le Gal ◽  
Florence Robert-Gangneux ◽  
Yann Pépino ◽  
Sorya Belaz ◽  
Céline Damiani ◽  
...  

Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 188
Author(s):  
Tanja Hoffmann ◽  
Andreas Hahn ◽  
Jaco J. Verweij ◽  
Gérard Leboulle ◽  
Olfert Landt ◽  
...  

This study aimed to assess standard and harsher nucleic acid extraction schemes for diagnostic helminth real-time PCR approaches from stool samples. A standard procedure for nucleic acid extraction from stool and a procedure including bead-beating as well as proteinase K digestion were compared with group-, genus-, and species-specific real-time PCR assays targeting helminths and nonhelminth pathogens in human stool samples. From 25 different in-house and commercial helminth real-time PCR assays applied to 77 stool samples comprising 67 historic samples and 10 external quality assessment scheme samples positively tested for helminths, higher numbers of positive test results were observed after bead-beating-based nucleic acid extraction for 5/25 (20%) real-time PCR assays irrespective of specificity issues. Lower cycle threshold values were observed for one real-time PCR assay after the standard extraction scheme, and for four assays after the bead-beating-based scheme. Agreement between real-time PCR results after both nucleic acid extraction strategies according to Cohen’s kappa ranged from poor to almost perfect for the different assays. Varying agreement was observed in eight nonhelminth real-time PCR assays applied to 67 historic stool samples. The study indicates highly variable effects of harsh nucleic acid extraction approaches depending on the real-time PCR assay used.


Sign in / Sign up

Export Citation Format

Share Document