PCR-based detection of <i>Mycobacterium avium</i> subsp. <i>paratuberculosis</i> infection in cattle in South Korea using fecal samples

Abstract

Mycobacterium avium subsp. paratuberculosis (MAP) is the causative agent of bovine paratuberculosis (PTB). The first step in the control of PTB is the identification and isolation of sub-clinical fecal shedders from the herd. In the current study, real-time and nested PCR targeting MAP-specific genetic elements (IS900 and ISMAP02) DNA isolated from fecal samples were used to detect MAP infection in cattle. Of the 1,562 fecal samples obtained from 37 herds, regardless of diarrhea, 35 samples tested positive in both IS900-targeted real-time and ISMAP02-targeted nested PCR. At the herd level, 12 of the 37 herds were found to be positive for MAP. Detection rates of the PCR tests were similar to those reported for ELISA-based methods. These results suggest that PCR can be used to detect sub-clinical fecal shedders of MAP.

Detection of Mycobacterium avium subspecies paratuberculosis excretion in bovine feces, using quantitative real time polymerase chain reaction (Q-PCR)

“…Quantification of amplicon copies by q-PCR in DNA samples taken from bovine feces positive to IS900. signs of the disease (Park et al, 2016;Fang et al, 2002). Previously, Khol et al (2010), had followed for 380 days a 2.5 year-old cow that had become naturally infected with Map, sampling feces on 9 occasions.…”

MicroRNA profiling in bovine serum according to the stage of Mycobacterium avium subsp. paratuberculosis infection

“…The number of samples in Group A (n = 4) was the same, and Groups B (n = 11), C (n = 28), and D (n = 46) added samples for validation by real-time PCR, including those used for sequencing. The mGITC/SC method [ 32 ] was used for DNA extraction from animal feces, and real-time PCR was conducted to detect a combination of IS900 and IS MAP 02 amplification [ 33 ]. Next, the results of the serum ELISA were obtained by using a commercial ELISA kit (IDEXX Laboratories, Inc., USA).…”

Genetic diversity of bovineMycobacterium aviumsubsp.paratuberculosisdiscriminated by IS1311PCR-REA, MIRU-VNTR, and MLSSR genotyping

“…During 2013 and 2014, detection of MAP by PCR was conducted by using fecal material collected from 37 herds across 6 provinces of Korea [ 20 ]. From a total of 1,562 fecal DNA samples, 35 were shown to be positive for MAP IS 900 and ISMap 02 .…”

“…In Korea, the national prevalence of PTB in individual cattle was estimated at 3.3% to 7.1% until 2010 [ 31 ]. Recently, 12 of 37 herds (32.4%) tested in Korea were found to be MAP-positive, as determined by performing polymerase chain reaction (PCR) of fecal DNA samples [ 20 ]. Despite the high herd-level prevalence, there have been no reports on the molecular characterization of MAP strains isolated from cattle in Korea.…”