MEASUREMENT OF IL-12 (P40, P35), IL-23P19 AND IFN-MRNA IN DUODENAL BIOPSIES OF CATS WITH IBD AND HEALTHY CONTROLS USING QUANTITATIVE REVERSE TRANSCRIPTASE POLYMERASE CHAIN REACTION (QRT-PCR)

2016 ◽  
Vol 17 (2) ◽  
pp. 25-33
Author(s):  
WALY E ◽  
BIOURGE V ◽  
Iain Peters ◽  
DAY J ◽  
STOKES R ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Healthy Controls ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Polymerase Chain ◽  
Duodenal Biopsies

scholarly journals AB0691 ANALYSIS OF SARS-COV-2 ANTIBODIES IN NON-COVID-19 PATIENTS: COMPARISON BETWEEN SYSTEMIC SCLEROSIS PATIENTS AND HEALTHY CONTROLS

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 1379.1-1379
Author(s):  
L. Giardullo ◽  
C. Rotondo ◽  
A. Corrado ◽  
N. Maruotti ◽  
R. Colia ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Systemic Sclerosis ◽  
Autoimmune Disease ◽  
Reverse Transcriptase ◽  
Real Time ◽  
Polymerase Chain Reaction Assay ◽  
Nuclear Antigen ◽  
Healthy Controls ◽  
Chain Reaction ◽  
Polymerase Chain

Background:Previous study evidenced a cross-reactivity between Sars-Cov-2 antibodies and autoimmune tissue antigen involved in connective tissue diseases, as nuclear antigen (NA), extractable nuclear antigen (ENA), histone and collagen (1). No study has been published about the titer of Sars-Cov-2 antibodies in non-infected patients with autoimmune disease.Objectives:To evaluate the titer of SARS-CoV-2 antibodies in non-COVID-19 patients and compare it between systemic sclerosis (SSc) patients and healthy controls (HC).Methods:A total of 58 patients with SSc (who fulfilled ACR/EULAR 2013 SSc classification criteria) and 18 HC were enrolled. Sera of all participants were collected, and SARS-CoV-2 antibodies (IgG and IgM) were evaluated by means ELISA. In all participants swabs for SARS-CoV-2 by real-time reverse-transcriptase-polymerase-chain-reaction assay were reported negative. Demographic, clinical, and autoimmune serological characteristics of SSc patients were recorded. The normal distribution was assessed using the Shapiro–Wilk’s test. Exclusion criteria was previous or actual Sars-Cov-2 infection. Comparisons between study groups of patients were evaluated by the Student’s t-test or Mann – Whitney U-test as appropriate. The differences between categorial variables were assessed by Pearson chi-square or Fisher’s exact test, as opportune. Statistical significance was set at p ≤ 0.05.Results:We observed significant differences between SSc patients and HC in serum levels of Sars-Cov-2 antibodies (IgG: 1,4±2,1 AU/ml vs 0,36±0,19 AU/ml respectively (p=0,001); and IgM: 2,5±3,1 AU/ml vs 0,8±0,7 AU/ml (p=0,022)). In 5 SSc patients was found titer of Sars-Cov-2 antibodies (IgG) exceeding the cut-off, but the control of swabs for SARS-CoV-2 by real-time reverse-transcriptase-polymerase-chain-reaction assay were negative. No significative differences in Sars-Cov-2 autoantibodies titer were found in subgroup of SSc patients with or without ILD or PAH, limited or diffuse skin subset, and different autoantibodies profile. Furthermore, antibodies titer was not associated with different drugs (steroid, methotrexate, mofetil-mycophenolate and bosentan) in use.Conclusion:A cross mimicking between Sars-Cov-2 antibodies and antinuclear antibodies or anti ENA could be hypothesized. Further studies are necessary to unravel the reliability of Sars-Cov-2 antibodies detection in autoimmune disease.References:[1]Vojdani, A., Vojdani, E., & Kharrazian, D. (2021). Reaction of human monoclonal antibodies to SARS-CoV-2 proteins with tissue antigens: Implications for autoimmune diseases. Frontiers in Immunology, 11, 3679Disclosure of Interests:None declared

scholarly journals Cattle rabies: the effect of clinical evolution, viral genetic lineage, and viral load on the severity of histological lesions

2020 ◽  
Vol 40 (4) ◽  
pp. 227-233
Author(s):  
Claudia S. Wisser ◽  
André Thaler Neto ◽  
Helena B.C.R. Batista ◽  
Enio Mori ◽  
Maria E.R. Chierato ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcriptase ◽  
Fluorescent Antibody ◽  
Inflammatory Lesion ◽  
Genetic Lineage ◽  
Fluorescent Antibody Technique ◽  
Chain Reaction ◽  
Antibody Technique ◽  
Qrt Pcr ◽  
Polymerase Chain

ABSTRACT: Our objective was the characterization and staging of histological lesions in different anatomical sites of the central nervous system (CNS) of rabid cattle. The severity of the lesions was compared with the clinical stages of the disease, the variants of viral isolates, and with the load of virus. Thirty-one spontaneously affected rabid cattle the state of Santa Catarina underwent clinical follow-up and were eventually necropsied. CNS tissues were sampled and submitted to direct fluorescent antibody technique (DFAT), immunohistochemistry (IHC), routine histopathology with hematoxylin and eosin stain (HE), reverse transcriptase polymerase chain reaction (RT-PCR), and polymerase chain reaction in quantitative reverse transcriptase in real time (qRT-PCR). Affected cattle were allotted in four groups according to their clinical stage when euthanized: G1, euthanized while standing; G2, euthanized when in sternal recumbence; G3, euthanized when in lateral recumbence; and G4, affected cattle with natural death. In order to evaluate the degree of severity of the lesions and the presence of Negri bodies (NBs), the brain was sectioned at 9 sites. Additionally, spinal cord and trigeminal ganglion sections were examined. The intensity of the lesions was graded as either absent, mild, moderate, or marked, and the presence or absence of the NBs was noted. Histological lesions were characterized by lymphocytic and monocytic meningoencephalitis with NBs in 28 cases. In all analyzed groups, intensities of histological lesions ranging from mild to severe were observed. Brain regions with the highest inflammatory lesion intensity were the medulla at the level of obex, followed by the colliculus and thalamus. NBs were observed in a higher percentage in the cerebellum, followed by medulla at the obex level, striatum complex, and frontal telencephalon. The duration of the clinical course of the disease did not influence the intensity of the inflammatory lesion, but it did influence the presence of NBs, with a higher percentage of these inclusions in cattle that died naturally than in euthanized cattle. All isolated rhabdovirus included in this study were genetically compatible with samples from hematophagous bats Desmodus rotundus. The evaluation by qRT-PCR did not demonstrate a correlation between lesion intensity and the amount of virus.

scholarly journals Rapid, Sensitive, and Specific Severe Acute Respiratory Syndrome Coronavirus 2 Detection: A Multicenter Comparison Between Standard Quantitative Reverse-Transcriptase Polymerase Chain Reaction and CRISPR-Based DETECTR

2020 ◽  
Author(s):  
Eelke Brandsma ◽  
Han J M P Verhagen ◽  
Thijs J W van de Laar ◽  
Eric C J Claas ◽  
Marion Cornelissen ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Severe Acute Respiratory Syndrome ◽  
Reverse Transcriptase ◽  
N Gene ◽  
Analytical Sensitivity ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Ribonucleoprotein Complexes ◽  
Large Patient ◽  
Polymerase Chain

Abstract Background Recent advances in CRISPR-based diagnostics suggest that DETECTR, a combination of reverse-transcriptase loop-mediated isothermal amplification (RT-LAMP) and subsequent Cas12 bystander nuclease activation by amplicon-targeting ribonucleoprotein complexes, could be a faster and cheaper alternative to quantitative reverse-transcription polymerase chain reaction (qRT-PCR) without sacrificing sensitivity and/or specificity. Methods In this study, we compare DETECTR with qRT-PCR to diagnose coronavirus disease 2019 on 378 patient samples. Patient sample dilution assays suggest a higher analytical sensitivity of DETECTR compared with qRT-PCR; however, this was not confirmed in this large patient cohort, where we report 95% reproducibility between the 2 tests. Results These data showed that both techniques are equally sensitive in detecting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) providing additional value of DETECTR to the currently used qRT-PCR platforms. For DETECTR, different guide ribonucleic acids can be used simultaneously to obviate negative results due to mutations in N-gene. Lateral flow strips, suitable as a point-of-care test, showed a 100% correlation to the high-throughput DETECTR assay. More importantly, DETECTR was 100% specific for SARS-CoV-2 relative to other human coronaviruses. Conclusions Because there is no need for specialized equipment, DETECTR could be rapidly implemented as a complementary technically independent approach to qRT-PCR thereby increasing the testing capacity of medical microbiological laboratories and relieving the existent PCR platforms for routine non-SARS-CoV-2 diagnostic testing.

scholarly journals Pendeteksian petanda kepuncaan glioblastoma multiforme

2020 ◽  
Vol 3 (1) ◽  
pp. 39-45
Author(s):  
Yohana Yohana
Keyword(s):  
Polymerase Chain Reaction ◽  
Glioblastoma Multiforme ◽  
Western Blot ◽  
Reverse Transcriptase ◽  
Real Time ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Polymerase Chain

Glioblastoma multiforme (GBM) adalah tumor otak ganas yang memiliki populasi sel punca kanker yang dapat  mempertahankan formasi tumor. Peranan sel punca kanker telah banyak dipelajari yang memiliki tanggung jawab terhadap resistensi dan rekurensi terapi GBM seperti radiasi dan kemoterapi. Beberapa petanda kepuncaan dapat dipakai diantaranya CD133, Nestin, A2B5, CD44, SOX2 and OCT4. Pemeriksaan histopatologi terhadap jaringan tumor yang dioperasi menjadi standar baku untuk menentukan derajat, tingkat keganasan, dan prognosis keganasan. Namun di sisi lain, kehadiran populasi sel punca yang memiliki sifat mampu memperbaharui diri dan mampu menginduksi pembentukan tumor memerlukan pemeriksaan yang lebih mendalam mengenai karakteristik biologi sel tumor. Pemeriksaan  sel punca dilakukan menggunakan flowcytometry dan imunohistokimia. Pemeriksaan petanda kepuncaan glioblastoma dilakukan dengan quantitative Reverse Transcriptase Real Time Polymerase Chain Reaction (qRT-PCR), Enzyme Linked Immunoabsorbent Assay (ELISA), Western Blot, Imunohistokimia dan flowcytometry. Petanda CD133 ditemukan ekspresinya meningkat pada berbagai pemeriksaan. CD133 digunakan sebagai prognostik GBM.

Breast Cancer Prognosis Determined by Gene Expression Profiling: A Quantitative Reverse Transcriptase Polymerase Chain Reaction Study

2005 ◽  
Vol 23 (29) ◽  
pp. 7278-7285 ◽  
Author(s):  
E. Espinosa ◽  
J.A. Fresno Vara ◽  
A. Redondo ◽  
J.J. Sánchez ◽  
D. Hardisson ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Polymerase Chain Reaction ◽  
Overall Survival ◽  
Lymph Node ◽  
Reverse Transcriptase ◽  
Gene Profile ◽  
Chain Reaction ◽  
Qrt Pcr ◽  
Polymerase Chain

Purpose We sought to reproduce with quantitative reverse-transcriptase polymerase chain reaction (qRT-PCR) the results obtained with a 70-gene expression profile that has been described previously in breast cancer. Patients and Methods Frozen breast cancer samples from patients who were operated on were used to isolate tumor RNA. Ninety-six patients with stage I to II disease were included. Median age was 57 years (range, 27 to 80 years). Forty-eight patients had lymph node–negative and 48 lymph node–positive disease. qRT-PCR amplifications were performed and the results were correlated with clinical data. Results After a minimum follow-up of 5 years, 25 patients had a relapse. The gene profile divided patients into two groups with poor and good prognosis. Significant differences with regard to grade of differentiation, size and hormone receptors were seen between the two groups. The gene profile was significantly associated with relapse-free survival and overall survival in the whole group of 96 patients. Multivariate analysis showed that only lymph node status and gene profile were significantly correlated to overall survival. Conclusion qRT-PCR reproduced the results obtained with microarrays for a prognostic gene profile in women with early-stage breast cancer.

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.

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