PCR Detection ofCoxiella burnetiifrom Different Clinical Specimens, Especially Bovine Milk, on the Basis of DNA Preparation with a Silica Matrix

Abstract

For PCR detection of Coxiella burnetii in various clinical specimens we developed a sample preparation method in which silica binding of DNA was used. This method was found to be fast, easily performed with large numbers of samples, and equally sensitive for all of the specimens tested (livers, spleens, placentas, heart valves, milk, blood). The DNA preparation method described here can also be used as an initial step in any PCR-based examination of specimens. The procedure was tested with more than 600 milk samples, which were taken from 21 cows that were seropositive for C. burnetii and reportedly had fertility problems (and therefore were suspected of shedding the agent through milk intermittently or continuously). Of the 21 cows tested, 6 were shedding C. burnetii through milk. Altogether, C. burnetii DNA was detected in 6% of the samples. There was no correlation between the shedding pattern and the serological results.

Molecular Detection of Rickettsia spp. and Coxiella burnetii in Cattle, Water Buffalo, and Rhipicephalus (Boophilus) microplus Ticks in Luzon Island of the Philippines

“…Coxiellosis can cause reproductive problems, such as metritis [27], abortion, delivery of premature offspring, stillbirth, and weak offspring (or APSW complex, as termed by Agerholm [56]) in animals. On another note, subclinically infected animals identified in this study present a greater risk to the people working with them, because they can shed C. burnetii through their feces, vaginal fluids, milk, and parturition byproducts [9,[57][58][59][60] without being identified as infected, due to absence of clinical signs. Moreover, six of those C. burnetii-positive animals are being raised for milk production, hence posing a health risk if the milk from those animals is consumed by humans without being pasteurized.…”

Extraction-Free, Filter-Based Template Preparation for Rapid and Sensitive PCR Detection of Pathogenic Parasitic Protozoa

“…These factors have significantly influenced the ability to reliably detect C. cayetanensis on raspberries. Steps to abrogate their effects have relied heavily on methods either to adsorb inhibitory substances from DNA extracts with polyvinyl polypyrrolidone (12,24), to bind DNA to a silica matrix in the presence of chaotropic reagents (12,26), or to dilute template preparations (22). While these steps, alone or in combination, have reduced PCR inhibition, a concomitant loss in detection signal has also been observed, and success has been marginal.…”

Scientific opinion on an alternative method for the hygienic treatment of bovine colostrum through a series of filtration steps

“…A comprehensive list of pathogens that could be present in colostrum was not provided by the applicant. The EFSA Panel on Biological Hazards (BIOHAZ Panel) considered the list of main hazards potentially present in bovine colostrum (those mentioned by the applicant are highlighted with an asterisk) (Peterson, 1965;Kawakami et al, 1966;Richardson, 1970;Ménard et al, 1983;Timoney et al, 1988;Watts, 1988;Lorenz et al, 1998;Waage et al, 1999;Pardo et al, 2001;Mukherjee et al, 2004;Izumi et al, 2006;Biesenkamp-Uhe et al, 2007;Barlow et al, 2008;Cervinkova et al, 2013;EFSA BIOHAZ Panel, 2015).…”