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.

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 Detection of lumpy skin disease virus in cattle using real-time polymerase chain reaction and serological diagnostic assays in different governorates in Egypt in 2017

2019 ◽  
Vol 12 (7) ◽  
pp. 1093-1100 ◽  
Author(s):  
Gamil Sayed Gamil Zeedan ◽  
Ayman Hamid Mahmoud ◽  
Abeer Mostafa Abdalhamed ◽  
Khaled Abd El-Hamid Abd El-Razik ◽  
Manal Hamdy Khafagi ◽  
...  
Keyword(s):  
Clinical Signs ◽  
Rt Pcr ◽  
Conventional Pcr ◽  
Serum Samples ◽  
Lumpy Skin Disease ◽  
Blood Samples ◽  
Diagnostic Assays ◽  
Cattle Serum ◽  
Skin Biopsies ◽  
Polymerase Chain

Background and Aim: Lumpy skin disease (LSD), is a highly infectious viral disease of cattle, caused by LSD virus (LSDV) which belongs to the genus Capripoxvirus of family Poxviridae. In the summer of 2017, skin lesions suggestive of LSD were observed in cattle at several governorates in Egypt. This study aimed to detect LSDV in cattle specimens using rapid serological and molecular diagnostic assays. Materials and Methods: A total of 46 skin biopsies and uncoagulated blood samples were collected from cattle with LSD suggestive clinical signs, as well as 290 coagulated whole blood samples from cattle without skin lesion in different governorates in Egypt during the summer of 2017. Skin biopsies were used for virus isolation from the chorioallantoic membrane of 11-day-old specific pathogen-free embryonated chicken eggs (SPF-ECEs). LSDV was identified using conventional polymerase chain reaction (PCR), real-time PCR (RT-PCR), and fluorescent antibody technique (FAT) with specific hyperimmune serum against LSDV. Cattle sera were examined using indirect FAT (IFAT) and indirect enzyme-linked immunosorbent assay (ELISA). Results: Skin nodules and sitfast lesions were significant clinical signs observed in all LSD suspect cattle. SPF-ECEs, from which positive isolations were made and it showed characteristic inflammatory and focal white pock lesions. The isolated viruses were identified as LSDV by FAT, conventional gel-based PCR, and RT-PCR. Among the skin biopsies and corresponding blood samples, LSDV-positive samples percentage were 39.13 and 36.95 by RT-PCR, followed 34.78 and 28.26 by conventional PCR and then 32.6 and 26.8 by FAT, respectively. The total positive percentage of LSDV antibody detected in cattle serum samples were 17.93 and 14.48 by indirect ELISA and IFAT. Conclusion: LSDV was detected and identified in skin biopsies and corresponding blood samples of naturally infected cattle, more LSDV-positive samples were detected by RT-PCR, followed by conventional PCR and then FAT. The indirect ELISA detected more antibody-positive samples than the IFAT from cattle serum samples. The RT-PCR assay is simple, sensitive, rapid, and reliable for the detection of LSDV in blood and skin nodule biopsies of suspected cattle.

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.

Polymerase chain reaction amplification and typing of rotavirus in environmental samples

1995 ◽  
Vol 31 (5-6) ◽  
pp. 371-374 ◽  
Author(s):  
R. Gajardo ◽  
R. M. Pintó ◽  
A. Bosch
Keyword(s):  
Polymerase Chain Reaction ◽  
Environmental Samples ◽  
Polymerase Chain Reaction Amplification ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Group A ◽  
Reaction Amplification ◽  
Stool Specimens ◽  
Polymerase Chain

A reverse transcription polymerase chain reaction (RT-PCR) assay is described that has been developed for the detection and serotyping of group A rotavirus in stool specimens and concentrated and non-concentrated sewage specimens.

scholarly journals Subarachnoid diverticulum associated with feline infectious peritonitis in a Siberian cat

2020 ◽  
Vol 6 (2) ◽  
pp. 205511692094147
Author(s):  
Christopher Hoey ◽  
George Nye ◽  
Angela Fadda ◽  
Janet Bradshaw ◽  
Emi N Barker
Keyword(s):  
Cervical Spinal Cord ◽  
Serum Biochemistry ◽  
Acute Onset ◽  
Neurological Deficits ◽  
Rt Pcr ◽  
Feline Infectious Peritonitis ◽  
Case Summary ◽  
Choroid Plexitis ◽  
The Brain ◽  
Feline Coronavirus

Case summary A 7-month-old Siberian cat was presented for investigation of acute onset multifocal neurological deficits. Neurological examination documented dull mental status and an ambulatory left hemiparesis. Serum biochemistry documented marked hyperglobulinaemia. MRI of the brain identified marked leptomeningeal contrast enhancement extending along the brainstem caudally to involve the cranial cervical spinal cord. MRI of the cervical spine further identified a subarachnoid diverticulum that extended from the level of the obex to the C2–C3 vertebrae. Cerebrospinal fluid quantitative RT-PCR was positive for the presence of feline coronavirus. Histopathology revealed pyogranulomatous meningitis and choroid plexitis, uveitis and nephritis. Relevance and novel information This article describes the first reported case of a subarachnoid diverticulum associated with feline infectious peritonitis.

scholarly journals Is it enough for COVID-19 screening test? Limitation of swab test and general characteristics of mild symptom patients

2021 ◽  
Vol 104 (2) ◽  
pp. 003685042110261
Author(s):  
Sungwoo Choi ◽  
Hyo Jeong Choi ◽  
Ho Jung Kim
Keyword(s):  
Screening Test ◽  
Community Treatment ◽  
Upper Respiratory Tract ◽  
Test Results ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Positive Rate ◽  
Upper Respiratory ◽  
Polymerase Chain ◽  
Discharge Criteria

The most common method for SARS-CoV-2 testing is throat or nasal swabbing by real-time reverse transcription polymerase chain reaction (RT-PCR) assay. In South Korea, drive-through swab test is used for screening system and community treatment centers (CTCs), which admit and treat confirmed COVID-19 patients with mild symptoms, are being used. This retrospective study was conducted on patients admitted to a CTC on March 6, 2020. A total of 313 patients were admitted. The nasal and throat swabs were collected from the upper respiratory tract, and a sputum test was performed to obtain lower respiratory samples. The positive rate of the first set of test, sputum test was higher than that of the swab test ( p = 0.011). In the second set of test, 1 week after the first ones, the rate of positive swab tests was relatively high ( p = 0.026). In the first set of test, 66 of 152 (43.4%) patients showed 24-h consecutive negative swab test results, when the sputum test results were considered together, that number fell to 29 patients (19.1%) ( p < 0.001). Also, in the second set of test, 63 of 164 (38.4%) patients met the discharge criteria only when the swab test was considered; that number fell to 30 (18.3%) when the sputum test results were also considered ( p < 0.001). Using the swab test alone is insufficient for screening test and discharge decision. Patients who may have positive result in the sputum test can be missed.

scholarly journals Monitoring Current-Season Spread of Potato virus Y in Potato Fields Using ELISA and Real-Time RT-PCR

Plant Disease ◽  
2013 ◽  
Vol 97 (5) ◽  
pp. 641-644 ◽  
Author(s):  
Manphool S. Fageria ◽  
Mathuresh Singh ◽  
Upeksha Nanayakkara ◽  
Yvan Pelletier ◽  
Xianzhou Nie ◽  
...  
Keyword(s):  
Real Time ◽  
Potato Virus ◽  
Potato Virus Y ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Pcr Assay ◽  
Current Season ◽  
Seed Market ◽  
Polymerase Chain ◽  
Time Of Harvest

The current-season spread of Potato virus Y (PVY) was investigated in New Brunswick, Canada, in 11 potato fields planted with six different cultivars in 2009 and 2010. In all, 100 plants selected from each field were monitored for current-season PVY infections using enzyme-linked immunosorbent assay (ELISA) and real-time reverse-transcription polymerase chain reaction (RT-PCR) assay. Average PVY incidence in fields increased from 0.6% in 2009 and 2% in 2010 in the leaves to 20.3% in 2009 and 21.9% in 2010 in the tubers at the time of harvest. In individual fields, PVY incidence in tubers reached as high as 37% in 2009 and 39% in 2010 at the time of harvest. Real-time RT-PCR assay detected more samples with PVY from leaves than did ELISA. A higher number of positive samples was also detected with real-time RT-PCR from growing tubers compared with the leaves collected from the same plant at the same sampling time. PVY incidence determined from the growing tubers showed a significant positive correlation with the PVY incidence of tubers after harvest. Preharvest testing provides another option to growers to either top-kill the crop immediately to secure the seed market when the PVY incidence is low or leave the tubers to develop further for table or processing purposes when incidence of PVY is high.

scholarly journals BAALC and ERG Expression in Egyptian Patients with Acute Myeloid Leukemia, Relation to Survival and Response to Treatment

2016 ◽  
Vol 4 (2) ◽  
pp. 264-270 ◽  
Author(s):  
Aml Soliman ◽  
Asmaa Abdel Aal ◽  
Reham Afify ◽  
Noha Ibrahim
Keyword(s):  
Polymerase Chain Reaction ◽  
Acute Myeloid Leukemia ◽  
Reverse Transcription ◽  
Myeloid Leukemia ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Polymerase Chain ◽  
Age And Sex ◽  
Acute Myeloid

AIM: Aim was to detect Brain and Acute Leukemia, Cytoplasmic (BAALC) and ETS-related gene (ERG) expression in patients with acute myeloid leukemia (AML) as well as to study their biologic and prognostic impact on the disease outcome and survival.PATIENTS AND METHODS: The current study was carried out on 44 patients with denovo acute myeloid leukemia, as well as 44 age and sex matched controls. The quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) assay was performed for estimation of BAALC and ERG expression.RESULTS: The current study was carried out on 44 patients with denovo acute myeloid leukemia, as well as 44 age and sex matched controls. The quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) assay was performed for estimation of BAALC and ERG expression. BAALC was expressed in 36 (81.82%) of AML cases versus 10 (22.72%) of the control group which was highly statistically significant (P < 0.001). While ERG was positive in 39(88.64%) of cases and 8(18.18 %) of controls and that was also highly statistically significant (P < 0.001).CONCLUSION: Further researches still needed to clarify the role of BAALC and ERG in the pathogenesis of leukemia and their importance as targets for treatment of AML.

scholarly journals Detection of Aspergillus species in bone marrow transplant patients

2010 ◽  
Vol 4 (08) ◽  
pp. 511-516 ◽  
Author(s):  
Parisa Badiee ◽  
Abdolvahab Alborzi
Keyword(s):  
Bone Marrow ◽  
Real Time ◽  
Bone Marrow Transplant ◽  
Real Time Pcr ◽  
Clinical Signs ◽  
Marrow Transplant ◽  
Aspergillus Species ◽  
Pcr Assay ◽  
Blood Samples ◽  
Transplant Patients

Introduction:  Invasive aspergillosis is a severe complication of cytotoxic chemotherapies and bone marrow transplantation (BMT). The aim of this study was to assess the utility of a real-time PCR assay for the early diagnosis of Aspergillus species in blood samples from BMT patients. Methodology: Blood specimens (n = 993) from patients (n = 82) scheduled for BMT were collected prior to transplant and for 100 days post transplantation.  The specimens were later tested using an Aspergillus-specific real-time PCR assay. Cultures of clinical samples, along with sonography and computerized tomographic scans, were performed as standard of care. Results: Aspergillus DNA was positive in 94 sequential blood samples from 13 patients with clinical and radiological signs of infection. Samples from three of these patients were PCR-positive for Aspergillus in the first week of admission, prior to transplantation. Four patients with aspergillosis were cured with antifungal agents and nine died. An additional 12 patients without clinical signs of infection were PCR-positive on one occasion each, while two patients with clinical signs of infection were PCR-negative. Compared to routine methods of aspergillosis diagnosis, the respective sensitivity, specificity, negative, and positive predictive values of the PCR method by patient were 86.6%, 82%, 96.5% and 52%. Conclusions: The results show that Aspergillus infections in the blood of bone marrow transplant patients can be dectected by PCR methods. Early detection of Aspergillus infections by PCR has the potential to positively impact patient mortality rate and provide cost savings to hospitals.

scholarly journals A beginner’s guide to RT-PCR, qPCR and RT-qPCR

2020 ◽  
Vol 42 (3) ◽  
pp. 48-53 ◽  
Author(s):  
Grace Adams
Keyword(s):  
Gene Expression ◽  
Polymerase Chain Reaction ◽  
Messenger Rna ◽  
Life Science ◽  
Science Research ◽  
Quantitative Real Time Pcr ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Mrna Quantitation ◽  
Polymerase Chain

The development of the polymerase chain reaction (PCR), for which Kary Mullis received the 1992 Novel Prize in Chemistry, revolutionized molecular biology. At around the time that prize was awarded, research was being carried out by Russel Higuchi which led to the discovery that PCR can be monitored using fluorescent probes, facilitating quantitative real-time PCR (qPCR). In addition, the earlier discovery of reverse transcriptase (in 1970) laid the groundwork for the development of RT-PCR (used in molecular cloning). The latter can be coupled to qPCR, termed RT-qPCR, allowing analysis of gene expression through messenger RNA (mRNA) quantitation. These techniques and their applications have transformed life science research and clinical diagnosis.

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