Immunohistochemical Methods to Diagnose Atraumatic Spleen Rupture in Feline Infectious Peritonitis of Tiger (Panthera tigris)

2017 ◽  
Vol 68 (5) ◽  
pp. 1055-1057
Author(s):  
Cristina Horhogea ◽  
Viorel Floristean ◽  
Mircea Lazar ◽  
Carmen Cretu ◽  
Carmen Solcan
Keyword(s):  
Peritoneal Fluid ◽  
Panthera Tigris ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Intestinal Lamina Propria ◽  
Feline Infectious Peritonitis ◽  
Liver And Kidney ◽  
Polymerase Chain ◽  
Immunohistochemical Methods ◽  
Feline Coronavirus

A body of an eight-year-old male tiger, originated from a Romanian zoo was brought in for pathological diagnosis to the Faculty of Veterinary Medicine. The necropsy revealed uveitis, sero-hemorrhagic peritonitis, fatty liver and kidney, haemorrhagic-necrotic lesions on the spleen, catarrhal enteritis, serous pericarditis and pulmonary anthracosis. Amyloidosis was found in the intestine, liver, lungs and kidney and spleen, associated with necrosis and the presence of hematin blocks. Positive macrophages for feline coronavirus (FCoV) were highlighted by immunohistochemistry in liver, kidney, lungs and intestinal lamina propria. Immunofluorescence (IF) examination of pericardial and peritoneal fluid was positive for feline infectious peritonitis (FIP). Confirmation of FIP coronavirus was made by Reverse Transcription Polymerase Chain Reaction (RT-PCR). The systemic amyloidosis and the presence of hematin in peritoneal fluid and spleen parenchyma are lesions that were not mentioned as specific for FIP until now.

scholarly journals Ontogeny of erythropoietin gene expression in the sheep fetus: effect of dexamethasone at 60 days of gestation

Blood ◽  
1994 ◽  
Vol 84 (2) ◽  
pp. 460-466
Author(s):  
GB Lim ◽  
K Jeyaseelan ◽  
EM Wintour
Keyword(s):  
Gene Expression ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Fetal Plasma ◽  
Liver And Kidney ◽  
Polymerase Chain ◽  
High Level ◽  
A Site ◽  
Sheep Fetus

We have used competitive reverse transcription and polymerase chain reaction (RT/PCR) to compare the levels of erythropoietin (Epo) mRNA in the liver and kidneys of the sheep fetus at 60, 80, 100, 130, and 140 days of gestation (term = 145 to 150 days). The effect of dexamethasone infusion in the ewe on Epo gene expression in the 60-day fetus was also investigated. Epo mRNA levels were highest at 60 days of gestation, the earliest age studied, in both liver and kidney. In the liver, Epo mRNA expression declined as gestation proceeded. Kidney Epo mRNA was maintained at a high level until 100 days of gestation, declining significantly in the 130-day fetus (P < .01). Treatment of ewes carrying 60-day fetuses with 0.76 mg/h dexamethasone for 48 hours resulted in a significant decrease in fetal plasma Epo values and Epo mRNA levels in both the liver and kidney. In the dexamethasone-treated fetuses, Epo mRNA in the liver was 52% of control values (P < .05), and in the kidney, 33% of control (P < .001). The results suggest that the kidney may play a more important role as a site of Epo synthesis in the early gestation sheep fetus than previously thought. Glucocorticoids may have a role in the regulation of Epo gene expression.

scholarly journals Comparison of real-time reverse transcriptase polymerase chain reaction of peripheral blood mononuclear cells, serum and cell-free body cavity effusion for the diagnosis of feline infectious peritonitis

2016 ◽  
Vol 19 (4) ◽  
pp. 344-350 ◽  
Author(s):  
Stephanie J Doenges ◽  
Karin Weber ◽  
Roswitha Dorsch ◽  
Robert Fux ◽  
Katrin Hartmann
Keyword(s):  
Real Time ◽  
Mononuclear Cells ◽  
Body Cavity ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Peripheral Blood Mononuclear ◽  
Feline Infectious Peritonitis ◽  
Blood Mononuclear Cells ◽  
Polymerase Chain ◽  
Free Body

Objectives Diagnosis of feline infectious peritonitis (FIP) remains challenging, especially in cats without effusions. The objective of this study was to evaluate the sensitivity and specificity of a real-time reverse transcriptase polymerase chain reaction (RT-PCR) detecting feline coronavirus (FCoV) RNA in peripheral blood mononuclear cells (PBMCs) and serum in comparison with the same real-time RT-PCR in cell-free body cavity effusion. Methods This prospective case-control study included 92 cats. Forty-three cats had a definitive diagnosis of FIP, established either by histopathological examination (n = 28) or by positive immunofluorescence staining of FCoV antigen in macrophages of effusions (n = 11), or by both methods (n = 4). Forty-nine control cats had other diseases but similar clinical signs. Real-time RT-PCR was performed on PBMCs of 37 cats (21 cats with FIP, 16 controls), on serum of 51 cats (26 cats with FIP, 25 controls) and on cell-free body cavity effusion of 69 cats (36 cats with FIP, 33 controls). Sensitivity, specificity, positive and negative predictive value, including 95% confidence intervals (CI), were calculated. Results Real-time RT-PCR of PBMCs, serum and cell-free body cavity effusion showed a specificity of 100% (95% CI 79.4–100% in PBMCs, 86.3–100% in serum, 89.4–100% in cell-free body cavity effusion) and a sensitivity of 28.6% (95% CI 11.3–52.2%) in PBMCs, 15.4% (95% CI 4.4–34.9%) in serum and 88.9% (95% CI 73.9–96.9%) in cell-free body cavity effusion to diagnose FIP. Conclusions and relevance Although it is known that RT-PCR can often provide false-positive results in healthy cats, this real-time RT-PCR was shown to be a specific tool for the diagnosis of FIP when applied in a clinical setting. Sensitivity in cell-free body cavity effusion was high but low in PBMCs and serum. PBMC samples showed a higher sensitivity than serum samples, and are therefore a better choice if no effusion is present.

scholarly journals Endothelin-1–Mediated Alteration of Metallothionein and Trace Metals in the Liver and Kidneys of Chronically Diabetic Rats

2002 ◽  
Vol 3 (3) ◽  
pp. 193-198 ◽  
Author(s):  
Lu Cai ◽  
Shali Chen ◽  
Terry Evans ◽  
M. George Cherian ◽  
Subrata Chakrabarti
Keyword(s):  
Trace Metals ◽  
Diabetic Rats ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Endothelin 1 ◽  
Liver And Kidney ◽  
Polymerase Chain ◽  
Age And Sex

In the present study, the role of endothelin-1 (ET-1) on alterations of hepatic and renal metallothionein (MT) and trace metals (Zn, Cu, and Fe) were investigated in streptozotocin (STZ)- induced diabetic rats. Diabetic rats, age- and sex-matched controls, as well as control and diabetic animals on a dualETA/ETBreceptor blocker, bosentan, were investigated after 6 months of follow-up. MT was measured by cadmium-heme assay. Metals were measured by atomic absorption spectrometer. ET-1 mRNA was analyzed by reverse transcriptase–polymerase chain reaction (RT-PCR) technique. Hepatic and renal ET-1 mRNA was increased in diabetic rats as compared to control rats, along with an increase in both hepatic and renal MT proteins. The increased hepatic MT protein level was associated with decreases in hepatic Cu and Fe, whereas increased renal MT was associated with increases in renal Cu and Fe accumulation. Zn levels were unaltered in both organs in diabetic rats. Bosentan treatment partially prevented the increase in MT levels in both liver and kidney, along with reduced serum creatinine and increased urinary creatinine levels. Further bosentan treatment corrected the increased Cu and Fe levels in the kidney in diabetic rats, but reduced hepatic Cu and Fe levels. No significant effects of bosentan treatment on nondiabetic rats were observed. The data suggest that the possible effects of ET antagonism in diabetes may be mediated via changes in MT and trace metals.

scholarly journals Ontogeny of erythropoietin gene expression in the sheep fetus: effect of dexamethasone at 60 days of gestation

Blood ◽  
1994 ◽  
Vol 84 (2) ◽  
pp. 460-466 ◽  
Author(s):  
GB Lim ◽  
K Jeyaseelan ◽  
EM Wintour
Keyword(s):  
Gene Expression ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Fetal Plasma ◽  
Liver And Kidney ◽  
Polymerase Chain ◽  
High Level ◽  
A Site ◽  
Sheep Fetus

Abstract We have used competitive reverse transcription and polymerase chain reaction (RT/PCR) to compare the levels of erythropoietin (Epo) mRNA in the liver and kidneys of the sheep fetus at 60, 80, 100, 130, and 140 days of gestation (term = 145 to 150 days). The effect of dexamethasone infusion in the ewe on Epo gene expression in the 60-day fetus was also investigated. Epo mRNA levels were highest at 60 days of gestation, the earliest age studied, in both liver and kidney. In the liver, Epo mRNA expression declined as gestation proceeded. Kidney Epo mRNA was maintained at a high level until 100 days of gestation, declining significantly in the 130-day fetus (P < .01). Treatment of ewes carrying 60-day fetuses with 0.76 mg/h dexamethasone for 48 hours resulted in a significant decrease in fetal plasma Epo values and Epo mRNA levels in both the liver and kidney. In the dexamethasone-treated fetuses, Epo mRNA in the liver was 52% of control values (P < .05), and in the kidney, 33% of control (P < .001). The results suggest that the kidney may play a more important role as a site of Epo synthesis in the early gestation sheep fetus than previously thought. Glucocorticoids may have a role in the regulation of Epo gene expression.

scholarly journals A comparative study of techniques used for the diagnosis of effusive feline infectious peritonitis

2020 ◽  
Vol 89 (2) ◽  
pp. 100-110
Author(s):  
A. Hellemans ◽  
D. D. Acar ◽  
V. J. E. Stroobants ◽  
S. Theuns ◽  
L. M. B. Desmarets ◽  
...  
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
High Sensitivity ◽  
Good Alternative ◽  
Chain Reaction ◽  
Reading Frame ◽  
Feline Infectious Peritonitis ◽  
Antibody Titers ◽  
Polymerase Chain ◽  
Low Sensitivity ◽  
Feline Coronavirus

Feline infectious peritonitis (FIP) is a fatal disease caused by feline infectious peritonitis virus (FIPV). At present, neither a licensed treatment nor an accurate ante-mortem diagnosis are available. In the present study, three available tests were evaluated for their diagnostic power on effusion samples. High feline coronavirus antibody titers, measured with an immunoperoxidase monolayer assay (IPMA), were correlated with FIP but its low specificity precluded a reliable diagnosis. The in-house 5’ reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) provided a much better specificity and high sensitivity. Given the low sensitivity of immunofluorescence staining (IF) of effusive cells, the RT-qPCR alone or in combination with IPMA represents a good alternative for IF. In the majority of the effusion samples from FIP positive animals, Sanger sequencing of the open reading frame encoding the spike protein (ORF S) revealed not only mutations that were previously associated with FIP (M1058L, S1060A, I1106T and D1108Y/E/G) but also two new, closely related mutations (T1112S/N).

scholarly journals Concordance between Histology, Immunohistochemistry, and RT-PCR in the Diagnosis of Feline Infectious Peritonitis

Pathogens ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 852
Author(s):  
Angelica Stranieri ◽  
Donatella Scavone ◽  
Saverio Paltrinieri ◽  
Alessia Giordano ◽  
Federico Bonsembiante ◽  
...  
Keyword(s):  
Accurate Information ◽  
Rt Pcr ◽  
Likelihood Ratios ◽  
Predictive Values ◽  
Feline Infectious Peritonitis ◽  
Mesenteric Lymph ◽  
Polymerase Chain ◽  
Sensitivity Specificity ◽  
Feline Coronavirus

Histology, immunohistochemistry (IHC), and reverse transcription polymerase chain reaction (RT-PCR) have been used to diagnose feline infectious peritonitis (FIP), but no information regarding the comparison of their diagnostic performances on the same organ is available. The aims of this study were to determine the concordance among these tests and to evaluate which combination of tests and organs can be used in vivo. Histology, IHC, and nested RT-PCR (RT-nPCR) for feline coronavirus (FCoV) were performed on spleen, liver, mesenteric lymph node, kidney, large and small intestine, and lung from 14 FIP and 12 non-FIP cats. Sensitivity, specificity, predictive values, likelihood ratios, and concordance were calculated. IHC and RT-nPCR had the highest concordance in lung and liver, histology and IHC in the other organs. The sensitivity of histology, IHC, and RT-nPCR on the different organs ranged from 41.7 to 76.9%, 46.2 to 76.9%, and 64.3 to 85.7%, respectively, and their specificity ranged from 83.3 to 100.0%, 100% and 83.3 to 100.0%. Therefore, IHC is recommended when histology is consistent with FIP. If RT-nPCR is performed as the first diagnostic approach, results should always be confirmed with IHC. Lung or liver provide accurate information regardless of the method, while IHC is preferred to RT-nPCR to confirm FIP in the kidney or intestine.

scholarly journals The detection of feline coronaviruses in blood samples from cats by mRNA RT-PCR

2007 ◽  
Vol 9 (5) ◽  
pp. 369-372 ◽  
Author(s):  
Kezban Can-Şahna ◽  
Veysel Soydal Ataseven ◽  
Dilek Pınar ◽  
Tuba Çiğdem Oğuzoğlu
Keyword(s):  
Messenger Rna ◽  
Clinical Signs ◽  
High Specificity ◽  
High Rate ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Blood Samples ◽  
Feline Infectious Peritonitis ◽  
Polymerase Chain ◽  
Feline Coronavirus

In this study, 26 blood samples were collected from 25 healthy cats and one cat with clinical signs suggestive of feline infectious peritonitis (FIP), namely, fever, weight loss, enlarged abdomen, and ascites. Blood samples were tested for feline coronavirus (FCoV) messenger RNA (mRNA) by an reverse transcriptase-polymerase chain reaction (RT-PCR) assay which has previously been described to have a high specificity in the diagnosis of clinical FIP [Simons AF, Vennema H, Rofina JE, Pol JM, Horzinek MC, Rottier PJM, Egberink HF (2005) A mRNA PCR for the diagnosis of feline infectious peritonitis. Journal of Virological Methods124, 111–116]. Overall we found 14 (54%) of the cats were positive for FCoV including the cat with clinical disease, but the high rate of positivity among healthy cats suggested a poor specificity for the clinical diagnosis of FIP among these cats. It was observed that the positivity rate was highest in cats aged between 6 months–1 year old. Our findings suggest that FCoVs may be present in the blood samples from healthy cats as well as cats with clinical FIP.

scholarly journals Feline coronavirus quantitative reverse transcriptase polymerase chain reaction on effusion samples in cats with and without feline infectious peritonitis

2016 ◽  
Vol 19 (2) ◽  
pp. 240-245 ◽  
Author(s):  
Louise Longstaff ◽  
Emily Porter ◽  
Victoria J Crossley ◽  
Sophie E Hayhow ◽  
Christopher R Helps ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Amino Acid ◽  
Reverse Transcriptase ◽  
Diagnostic Marker ◽  
Spike Protein ◽  
Chain Reaction ◽  
Feline Infectious Peritonitis ◽  
Qrt Pcr ◽  
Polymerase Chain ◽  
Feline Coronavirus

Objectives The aim of the study was to determine whether feline coronavirus (FCoV) RNA in effusion samples can be used as a diagnostic marker of feline infectious peritonitis (FIP); and in FCoV RNA-positive samples to examine amino acid codons in the FCoV spike protein at positions 1058 and 1060 where leucine and alanine, respectively, have been associated with systemic or virulent (FIP) FCoV infection. Methods Total RNA was extracted from effusion samples from 20 cats with confirmed FIP and 23 cats with other diseases. Feline coronavirus RNA was detected using a reverse transcriptase quantitative polymerase chain reaction assay (qRT-PCR), and positive samples underwent pyrosequencing of position 1058 with or without Sanger sequencing of position 1060 in the FCoV spike protein. Results Seventeen (85%) of the effusion samples from 20 cats with FIP were positive for FCoV RNA, whereas none of the 23 cats with other diseases were positive. Pyrosequencing of the 17 FCoV-positive samples showed that 11 (65%) of the cats had leucine and two (12%) had methionine at position 1058. Of the latter two samples with methionine, one had alanine at position 1060. Conclusions and relevance A positive FCoV qRT-PCR result on effusions appears specific for FIP and may be a useful diagnostic marker for FIP in cats with effusions. The majority of FCoVs contained amino acid changes previously associated with systemic spread or virulence (FIP) of the virus.

1504: Molecular Diagnosis and Staging of Prostate Cancer Using Clusterin in Real Time Reverse Transcription Polymerase Chain Reaction (RT-PCR)

2006 ◽  
Vol 175 (4S) ◽  
pp. 485-486
Author(s):  
Sabarinath B. Nair ◽  
Christodoulos Pipinikas ◽  
Roger Kirby ◽  
Nick Carter ◽  
Christiane Fenske
Keyword(s):  
Prostate Cancer ◽  
Polymerase Chain Reaction ◽  
Real Time ◽  
Molecular Diagnosis ◽  
Reverse Transcription ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain
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