Comparison of PCR and histopathology methods for diagnosis of Yersiniosis infection (yersinia ruckeri) in Rainbow trout of Iran

Subject Areas : Journal of Comparative Pathobiology

عادل Haghighi Khiabanian Asl ¹ , M.R Roozbahani ² , بهرام Kazemi ³

1 - ندارد
2 - ندارد
3 - ندارد

Received: 2010-07-03 Accepted : 2010-10-07 Published : 2010-02-20

Keywords: Histopathology-Multiplex PCR-E,

Abstract :

Yersinia ruckeri is the causative agent of Yersiniosis or Enteric Red mouth Disease that causeshuge amount of economic loss in mariculture industry worldwide. Definitive detection andeffective treatment for septicemia like Y. ruckeri that outbreak critically, requires rapid andspecific diagnosis. Despite the advantages of both PCR and Histopathology techniques in theterms of diagnosis and monitoring they have some defects, the aim of this study is thecomparison of PCR and histopathology methods for detection of Yersiniosis in trout fish inIran.In PCR-based detection, Extracted DNA from suspected Rainbow trout tissue utilized inPCR reaction and polymerase chain products were visualized by gel electrophoresis. Inpathological detection, all tissues from live or moribund fish were fixed in 10% formalin saline.Then Samples were embedded in paraffin block and sectioned with digital microtome at 5-7micrometer thickness. Slides were stained by haematoxillin & eosin, and then treated withmounting media. Differences in the number of positive samples in this comparison demonstratedthat each mentioned technique has its advantages so if the advantages of each method exploitand one method supplements the other one, the efficiency and accuracy of monitoring theexperiments would boost. In both mentioned methods, all positive results confirmed theexistence of yersiniosis infection in Iran, which their comparison is very useful for monitoringand diagnosis of pathogen in the country. Differences in the number of positive samples in thiscomparison demonstrate that although PCR technique would provide a more rapid and sensitivealternative to traditional diagnosis in detection of harmful pathogens but in some cases such asnecrotic and degenerated cells in tissue, PCR ability collapses in the comparison of otherdetection technique like histopathology. In this study, 20 suspected samples were tested by bothtechniques that 2 samples were positive and 18 samples were negative in PCR, and 5 sampleswere positive and 15 samples were negative in pathologic technique .