Composite testing for ante-mortem diagnosis of Johne’s disease in farmed New Zealand deer: correlations between bacteriological culture, histopathology, serological reactivity and faecal shedding as determined by quantitative PCR

BMC Veterinary Research

Frank Griffin¹  Simon Liggett¹  Alan Hughes¹  Rory O’Brien¹ 

[1] Disease Research Laboratory, Department of Microbiology & Immunology, University of Otago, Dunedin, New Zealand

关键词: Cervus elaphus;    Deer;    ELISA;    Quantitative PCR;    MAP;    Paratuberculosis;    Johne’s disease;   

Others  :  1119566
DOI  :  10.1186/1746-6148-9-72

 received in 2013-02-05, accepted in 2013-04-01,  发布年份 2013

【 摘 要 】

Background

In the absence of overt clinical signs of Johne’s Disease (JD), laboratory based tests have largely been limited to organism detection via faecal culture or PCR and serological tests for antibody reactivity. In this study we describe the application of quantitative faecal PCR for the detection of Mycobacterium avium subsp. paratuberculosis (MAP) in New Zealand farmed deer to quantify the bacterial load in cervine faecal samples as an adjunct to an existing serodiagnostic test (Paralisa™) tailored for JD diagnosis in deer. As ELISA has potential as a cheap, high throughput screening test for JD, an attempt was made to assess the sensitivity, specificity and positive/negative predictive (PPV/NPV) values of Paralisa™ for estimating levels of faecal shedding of MAP as a basis for JD management in deer.

Results

Correlations were made between diagnostic tests (ELISA, qPCR, culture and histopathology) to establish the precision and predictive values of individual tests. The findings from this study suggest there is strong correlation between bacterial shedding, as determined by faecal qPCR, with both culture (r = 0.9325) and histopathological lesion severity scoring (r = 0.7345). Correlation between faecal shedding and ELISA reactivity in deer was weaker with values of r = 0.4325 and r = 0.4006 for Johnin and Protoplasmic antigens, respectively. At an ELISA Unit (EU) cutoff of >50 (Johnin antigen) the PPV of Paralisa™ for significant faecal shedding in deer (>10⁴ organisms/g) was moderate (0.55) while the NPV was higher (0.89). At an EU cutoff of ≥150, the PPV for shedding >10⁵ organisms/g rose to 0.88, with a corresponding NPV of 0.85.

Conclusions

The evidence available from this study suggests that Paralisa™ used at a cutoff of 50EU could be used to screen deer herds for MAP infection with sequential qPCR testing used to cull all Paralisa™ positive animals that exhibit significant MAP faecal shedding.

【 授权许可】

   
2013 O’Brien et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

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