Comparison of C ₁₈ -Carboxypropylbetaine and Glass Bead DNA Extraction Methods for Detection of Mycobacterium bovis in Bovine Milk Samples and Analysis of Samples by PCR

Abstract

The purpose of this prospective study was to compare two different milk preparation methods to assay for the presence ofMycobacterium bovis by PCR. Detection by a C18-carboxypropylbetaine (CB-18)-based sample processing method was compared to extraction of DNA from milk with glass beads. Samples from 17 skin test-positive cattle were analyzed. Following CB-18 processing and glass bead extraction, the sensitivity of IS6110-based PCR was 94.1 and 58.8%, respectively (P < 0.025). Because CB-18 processing will permit the proficient use of PCR for diagnosis and surveillance of bovine tuberculosis, it will contribute to the more efficient detection and control of tuberculosis.

Individual Animals of a Cattle Herd Infected with the Same Mycobacterium bovis Genotype Shows Important Variations in Bacteriological, Histopathological and Immune Response Parameters

“…Importantly, a high number of animals present M. bovis in their noses, as demonstrated by PCR and culture. As it has already been observed with culture and PCR of M. bovis in bovine milk (Cornejo et al, 1998;Zumárraga et al, 2005), PCR is more sensitive than culture, a fact that may be attributable to the decontamination method before culture that may kill a high proportion of bacteria.…”

“…In conclusion, in a herd infected by the same strain, ante-mortem direct and immune-diagnostic parameters change, suggesting that several tests are needed for a faster control of infection at herd level. detect members of the M. tuberculosis complex and is especially useful for the direct detection of M. bovis in bovine tissue samples (Kolk et al, 1992;Kox et al, 1994;Liebana et al, 1995;Wards et al, 1995;Cornejo et al, 1998;Vitale et al, 1998;Zanini et al, 1998Zanini et al, , 2001Romero et al, 1999;Zumárraga et al, 2001Zumárraga et al, , 2005) Eradication of this disease remains an important goal in several countries. The strategy of test and slaughter has been used widely in an attempt to control dissemination of the disease and is based on the tuberculin skin test as a means for BTB diagnosis.…”

Evaluation of culture media for the recovery of Mycobacterium avium subsp. paratuberculosis from Cheddar cheese

“…Recovery of MAP from liquid milk relies on chemical decontamination of the sample to inactivate undesirable micro-organisms that could contaminate agar slopes. Although a range of decontamination procedures have been used (Sweeney et al 1992;Miller et al 1996;Cornejo et al 1998;Rahn et al 1998), Dundee et al (2001 concluded that the decontamination protocol involving 0AE75% (w/v) hexadecylpyridinium chloride (HPC) for 5 h at room temperature was the most sensitive for the recovery of MAP from liquid milk. While a decontamination step may be used to isolate MAP from cheese or other dairy products, such a step has implications for the accurate enumeration of MAP .…”

Comparative evaluation of four decontamination protocols for the isolation of Mycobacterium avium subsp. paratuberculosis from milk

“…paratuberculosis from milk and the method showed promise. The use of CB-18 in decontamination protocols for M. bovis has previously been reported (Cornejo et al 1998;Thornton et al 1998). CB-18 is a zwitterionic detergent and its use during decontamination may increase the recovery of mycobacteria by centrifugation (Anonymous 1998).…”

“…There is some uncertainty about whether current pasteurization conditions are capable of inactivating M.a. paratuberculosis if present in raw milk (Chiodini and Hermon-Taylor 1993;Grant et al 1996Grant et al , 1998Hope et al 1997;Stabel et al 1997). As a consequence, there has been recent increased activity towards developing and optimizing methods for the isolation of viable M.a.…”