scholarly journals Immunohistochemistry and Polymerase Chain Reaction for the Detection of Lawsonia intracellularis in Porcine Intestinal Tissues with Proliferative Enteropathy.

2000 ◽  
Vol 62 (7) ◽  
pp. 771-773 ◽  
Author(s):  
Junghyun KIM ◽  
Changsun CHOI ◽  
Wan-Seob CHO ◽  
Chanhee CHAE
2003 ◽  
Vol 15 (3) ◽  
pp. 268-273 ◽  
Author(s):  
Magdalena Jacobson ◽  
Stina Englund ◽  
András Ballagi-Pordány

Lawsonia intracellularis is an intracellular organism that causes proliferative enteritis in pigs. This bacterium is difficult to culture, and antemortem demonstration of the microbe is therefore often performed on fecal samples by polymerase chain reaction (PCR). Polymerase chain reaction is sensitive and specific, but inhibitory factors in feces might cause false-negative results. This article describes the construction and use of an internal standard, a mimic. The mimic is amplified by the same primers as those used for L. intracellularis DNA and thus could indicate false-negative results in clinical samples. The amplicon was clearly visible when as few as 10 mimic molecules were added per amplification reaction and when no inhibitors were present. When fecal samples were spiked with the mimic, the detection limit was 102 molecules per PCR. Sixty clinical samples, 20 from wild boars, 20 from growing pigs with diarrhea, and 20 from pigs without diarrhea, were prepared by a boiling procedure and subjected to PCR together with 103 mimic molecules. Nine samples were positive, of which 7 originated from pigs with diarrhea and 2 from pigs without diarrhea. In 14 samples from wild boars, in 8 samples from pigs without diarrhea, and in 3 samples from pigs with diarrhea, neither the mimic nor the target DNA was visible. This indicated the presence of inhibitors in these samples. It is concluded that the mimic can be used as an internal control in the diagnosis of L. intracellularis to indicate inhibition of PCR.


1994 ◽  
Vol 41 (3) ◽  
pp. 205-212 ◽  
Author(s):  
Steven McOrist ◽  
Connie J. Gebhart ◽  
Gordon H.K. Lawson

2021 ◽  
Vol 42 (5) ◽  
pp. 2867-2876
Author(s):  
Tatiane Caleffo ◽  
◽  
Vinicius Dahm ◽  
Jéssica Gonçalves dos Santos ◽  
Arthur Colombari Cheng ◽  
...  

Lawsonia intracellularis is a bacterium already described in several species and most prevalent in pigs, in which it causes enteric problems. Horses can also be affected, developing a disease known as equine proliferative enteropathy, which results from the proliferation of intestinal crypt cells in response to infection by the bacterium. Despite the existence of reports of the disease in several countries, including Brazil, there are still no reports of the disease or epidemiological studies of its occurrence in symptomatic or asymptomatic horses in the state of Paraná. Thus, the present study was conducted to examine the occurrence of L. intracellularis in asymptomatic horses raised in the west, northwest and north regions of Paraná by means of serological testing and the real-time polymerase chain reaction (qPCR) technique. In the serological approach, the immunoperoxidase monolayer assay (IPMA) technique was employed. Feces were processed and subjected to qPCR. In total, samples were collected from 162 animals from 20 farms. Of these, 9/162 (5.55%) showed specific antibodies against L. intracellularis. Real-time PCR, on the other hand, identified 7/162 (4.32%) fecal samples positive for the presence of the bacterium. When the techniques were compared, none of the samples was positive by both, demonstrating that, for a better diagnosis, they must be performed together. In contrast to most reports in horses, the present study describes higher serological and molecular occurrence in animals older than two years. These results are of great epidemiological relevance, as they indicate that the bacterium is present in the sampled regions of the state of Paraná. Therefore, the disease must be included in the differential diagnosis of diseases with similar clinical manifestations.


2014 ◽  
Vol 26 (3) ◽  
pp. 342-345 ◽  
Author(s):  
Ken Steen Pedersen ◽  
Markku Johansen ◽  
Sven Erik Jorsal ◽  
Jens Peter Nielsen ◽  
Poul Bækbo ◽  
...  

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