scholarly journals Rapid differentiation of Mycobacterium avium subsp. avium and Mycobacterium avium subsp. paratuberculosis by amplification of insertion element IS901

2012 ◽  
Vol 47 (No. 5) ◽  
pp. 117-121 ◽  
Author(s):  
P. Svastova ◽  
I. Pavlik ◽  
M. Bartos
Keyword(s):  
Mycobacterium Avium ◽  
False Positive ◽  
Mycobacterium Avium Complex ◽  
Mycobacterium Avium Subsp ◽  
Size Difference ◽  
Insertion Element ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Positive Results ◽  
Specific Amplicon

The aim of this study was to examine the specificity of primers designed to detect the insertion element IS901 commonly used in differentiation of Mycobacterium avium complex strains. This study shows that one of these primers non-specifically anneals to a sequence inside insertion element IS900, specific IS of M. avium subsp. paratuberculosis and to another sequence flanking this element. The resulting non-specific amplicon can be a product of amplification from some M. avium subsp. paratuberculosis strains and can simulate the presence of insertion element IS901 in these strains. However size difference between specific and non-specific amplicons allows such false-positive results to be distinguished. In addition the single PCR allows a rapid and simple differentiation between IS901+ M. avium subsp. avium and M. avium subsp. paratuberculosis strains.

scholarly journals Detection of Mycobacterium avium subsp. paratuberculosis in liver of slaughtered cattle, sheep and goats by PCR and culture

2021 ◽  
Vol 72 (1) ◽  
pp. 2683
Author(s):  
M ZAREI ◽  
S TAJBAKHSH ◽  
M GHORBANPOUR ◽  
A YOUSEFVAND ◽  
S RAHIMI ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Disease Status ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Mycobacterium Avium Subsp ◽  
Health Concern ◽  
Sheep And Goats ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Positive Results ◽  
Potential Public Health

The presence of Mycobacterium avium subsp. paratuberculosis (MAP) in tissues other than intestines and associated lymph nodes is a potential public health concern. Therefore, the presence of MAP in the liver of slaughtered cattle, sheep and goats with unknown Johne’s disease status was evaluated. Liver samples were collected randomly from 470 mature animals including 200 cattle, 135 sheep and 135 goats. The existence of MAP DNA from obtained liver samples was determined using PCR, thereafter PCR-confirmed samples were cultured. Based on liver PCR results, IS900 genes were detected in 11/200 (5.5%), 3/135 (2.2%), 2/135 (1.48%) liver samples of cattle, sheep and goats, respectively. In cattle, sheep and goats, 6 (54.5%), 1 (33.3%) and 1 (50%) PCR-positive samples showed positive results in culture, respectively. Our results revealed that MAP can be detected and cultured from the liver of slaughtered cattle, sheep and goats with unknown Johne’s disease status. Hence, given concerns about a potential zoonotic role for MAP, there is a crucial need to detect animals with MAP disseminated infection in the liver before slaughter.

scholarly journals Isolation of Mycobacterium avium Subsp. Paratuberculosis in the Feces and Tissue of Small Ruminants Using a Non-Automated Liquid Culture Method

Animals ◽  
2019 ◽  
Vol 10 (1) ◽  
pp. 20
Author(s):  
Luigi De Grossi ◽  
Davide Santori ◽  
Antonino Barone ◽  
Silvia Abbruzzese ◽  
Matteo Ricchi ◽  
...  
Keyword(s):  
Mycobacterium Avium ◽  
Liquid Culture ◽  
Culture Media ◽  
Small Ruminants ◽  
Culture Method ◽  
Mycobacterium Avium Subsp ◽  
Type I ◽  
Culture Methods ◽  
Specific Pcr ◽  
Mycobacterium Avium Subsp Paratuberculosis

Paratuberculosis is a chronic disease of ruminants caused by Mycobacterium avium subsp. Paratuberculosis (MAP). Since isolation of MAP type I (S) is rarely reported in Italy, our research was aimed at isolating, by an inexpensive liquid culture manual method, this type of MAP isolates. At first, we used an ELISA to point out to serologically positive samples from five flocks. Secondly, we used a fecal direct IS900-qPCR on the ELISA positive samples, in order to detect shedder animals. Feces from IS900-qPCR positive samples were inoculated in solid and liquid culture media. IS900-qPCR was further used to test the growth of MAP isolates in liquid medium, which were further confirmed by f57-qPCR and submitted to typing by specific PCR in order to identify the MAP type. Twenty-eight samples (24 fecal and four tissutal samples) were processed by culture methods, resulting in the isolation of six type I MAP field isolates. Notably, no isolates were recovered by solid media, underlining the utility of this liquid method. Few data about this type of MAP are currently available in Italy, and further analyses should be carried out in order to study the origin and epidemiology of type I strains circulating in Italy.

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