Rift Valley Fever Viral RNA Detection by In Situ Hybridization in Formalin-Fixed, Paraffin-Embedded Tissues

2019 ◽  
Vol 19 (7) ◽  
pp. 553-556 ◽  
Author(s):  
Izabela K. Ragan ◽  
Kaitlynn N. Schuck ◽  
Deepa Upreti ◽  
Lieza Odendaal ◽  
Juergen A. Richt ◽  
...  
2020 ◽  
Vol 32 (4) ◽  
pp. 577-580
Author(s):  
Brian J. Shiell ◽  
Siying Ye ◽  
Jennifer A. Harper ◽  
Brenda van der Heide ◽  
Gary Beddome ◽  
...  

Rift Valley fever virus (RVFV) causes Rift Valley fever (RVF), resulting in morbidity and mortality in humans and ruminants. Evidence of transboundary outbreaks means that RVFV remains a threat to human health and livestock industries in countries that are free from the disease. To enhance surveillance capability, methods for detection of RVFV are required. The generation of reagents suitable for the detection of RVFV antigen in formalin-fixed, paraffin-embedded tissues from infected animals have been developed and are described herein. Recombinant nucleoprotein (rNP) was expressed in Escherichia coli and purified using immobilized metal ion affinity chromatography. Purified rNP was used as an immunogen to produce anti-NP polyclonal antisera in rabbits for use in detection of RVFV NP in experimentally infected animals by immunohistochemistry. Antisera raised in rabbits against rNP were able to recognize viral NP antigen in fixed infected Vero cell pellets and sheep liver. Therefore, the methods and reagents described herein are useful in assays for detection of RVFV infections in animals, for research and surveillance purposes.


2018 ◽  
Vol 54 (1) ◽  
pp. 65-70 ◽  
Author(s):  
Joon Im ◽  
Derek P. Burney ◽  
Sean P. McDonough ◽  
Brigid Nicholson ◽  
Adam Eatroff ◽  
...  

ABSTRACT This case report describes the detection of intrahepatic bacteria in formalin-fixed paraffin-embedded histopathological sections from three dogs with neutrophilic, pyogranulomatous, or lymphoplasmacytic hepatitis and cholangiohepatitis. In each of these cases, eubacterial fluorescence in situ hybridization enabled colocalization of intrahepatic bacteria with neutrophilic and granulomatous inflammation in samples that were negative for bacteria when evaluated by routine hematoxylin and eosin histopathology augmented with histochemical stains. Positive responses to antimicrobial therapy were observed in of 2 out of 2 patients that were treated with antimicrobials. These findings suggest that eubacterial fluorescence in situ hybridization analysis of formalin-fixed paraffin-embedded histopathological sections is more sensitive than conventional histochemical stains for the diagnosis of bacteria-associated canine hepatitis.


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