scholarly journals Canine distemper virus detection by different methods of One-Step RT-qPCR

2016 ◽  
Vol 46 (9) ◽  
pp. 1601-1606
Author(s):  
Claudia de Camargo Tozato ◽  
Vívian Ferreira Zadra ◽  
Caroline Rodrigues Basso ◽  
João Pessoa Araújo Junior
Keyword(s):  
Urine Sample ◽  
Canine Distemper Virus ◽  
Limit Of Detection ◽  
Analytical Sensitivity ◽  
Canine Distemper ◽  
System A ◽  
Virus Rna ◽  
One Step ◽  
Commercial Kits ◽  
The One

ABSTRACT: Three commercial kits of One-Step RT-qPCR were evaluated for the molecular diagnosis of Canine Distemper Virus. Using the kit that showed better performance, two systems of Real-time RT-PCR (RT-qPCR) assays were tested and compared for analytical sensitivity to Canine Distemper Virus RNA detection: a One-Step RT-qPCR (system A) and a One-Step RT-qPCR combined with NESTED-qPCR (system B). Limits of detection for both systems were determined using a serial dilution of Canine Distemper Virus synthetic RNA or a positive urine sample. In addition, the same urine sample was tested using samples with prior centrifugation or ultracentrifugation. Commercial kits of One-Step RT-qPCR assays detected canine distemper virus RNA in 10 (100%) urine samples from symptomatic animals tested. The One-Step RT-qPCR kit that showed better results was used to evaluate the analytical sensitivity of the A and B systems. Limit of detection using synthetic RNA for the system A was 11 RNA copies µL-1 and 110 RNA copies µl-1 for first round System B. The second round of the NESTED-qPCR for System B had a limit of detection of 11 copies µl-1. Relationship between Ct values and RNA concentration was linear. The RNA extracted from the urine dilutions was detected in dilutions of 10-3 and10-2 by System A and B respectively. Urine centrifugation increased the analytical sensitivity of the test and proved to be useful for routine diagnostics. The One-Step RT-qPCR is a fast, sensitive and specific method for canine distemper routine diagnosis and research projects that require sensitive and quantitative methodology.

scholarly journals A New Molecular Detection System for Canine Distemper Virus Based on a Double-Check Strategy

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1632
Author(s):  
Sabrina Halecker ◽  
Sabine Bock ◽  
Martin Beer ◽  
Bernd Hoffmann
Keyword(s):  
Canine Distemper Virus ◽  
Parallel Implementation ◽  
Detection System ◽  
Quantitative Polymerase Chain Reaction ◽  
Diagnostic Methods ◽  
Molecular Diagnostic ◽  
Wild Animals ◽  
Canine Distemper ◽  
One Step ◽  
Diagnostic Sensitivity And Specificity

Due to changing distemper issues worldwide and to inadequate results of an inter-laboratory study in Germany, it seems sensible to adapt and optimize the diagnostic methods for the detection of the canine distemper virus (CDV) to the new genetic diversity of virus strains. The goal of the project was the development, establishment and validation of two independent one-step reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) methods for the safe detection of CDV in domestic and wild animals. For this purpose, an existing CDV-RT-qPCR was decisively adapted and, in addition, a completely new system was developed. Both CDV-RT-qPCR systems are characterized by a very high, comparable analytical and diagnostic sensitivity and specificity and can be mutually combined with inhibition or extraction controls. The reduction in the master mix used allows for the parallel implementation of both CDV-RT-qPCR systems without significant cost increases. For validation of the new CDV-RT-qPCR duplex assays, a panel comprising 378 samples derived from Germany, several European countries and one African country were tested. A sensitivity of 98.9% and a specificity of 100% were computed for the new assays, thus being a reliable molecular diagnostic tool for the detection of CDV in domestic and wild animals.

scholarly journals Studi Kasus: Paralisis pada Anjing Shih-tzu yang Diduga Terinfeksi Virus Distemper Anjing

2019 ◽  
pp. 34
Author(s):  
Santri Devita Sari Gurning ◽  
Sri Kayati Widyastuti ◽  
I Gede Soma
Keyword(s):  
Canine Distemper Virus ◽  
Canine Distemper ◽  
Virus Rna

Canine Distemper Virus (CDV) merupakan virus RNA beramplop, genus Morbilivirus dari famili Paramyxoviridae. Anjing Shih-tzu yang dijadikan kasus berumur dua tahun, dilaporkan mengalami kelumpuhan sejak dua bulan yang lalu. Anjing pernah didiagnosis CDV dengan gejala letargi, anoreksia, demam, batuk, keluar eksudat berlebih dari hidung dan mata. Anjing tahan terhadap infeksi awal CDV, dan beberapa minggu setelah infeksi, anjing menunjukkan gejala saraf. Berdasarkan riwayat penyakit, tanda klinis dan pemeriksaan darah anjing kasus terindikasi mengalami leukopenia, limfopenia, dan anemia, hewan kasus didiagnosis mengalami paralisis akibat ikutan penyakit distemper anjing. Terapi yang diberikan berupa terapi simptomatis dan supportif dengan pemberian infus NaCl fisiologis sebanyak 870 ml/hari selama 3 hari, injeksi Neurotropic® dengan dosis 0,5 ml sekali pemberian, injeksi Amoxycillin 10% dosis 0,5 ml sekali pemberian, serta antibiotik tabur (Enbatic®) yang ditaburkan pada kulit yang mengalami ulserasi sekali sehari sampai sembuh. Anjing dievaluasi tujuh hari kemudian dan tidak ditemukan tanda-tanda pemulihan dari gejala saraf namun kulit yang mengalami ulserasi terlihat membaik. Evaluasi dihari ke-14 juga tidak banyak perubahan, bekas ulserasi kulit yang tampak memudar. Pada hari ke-28 hewan beberapa kali mengalami epistaksis kemudian mati di hari berikutnya.

scholarly journals A fast and simple one-step duplex PCR assay for canine distemper virus (CDV) and canine coronavirus (CCoV) detection

2018 ◽  
Vol 163 (12) ◽  
pp. 3345-3349 ◽  
Author(s):  
Jing Wang ◽  
Yakun Luo ◽  
Lin Liang ◽  
Jinxiang Li ◽  
Shangjin Cui
Keyword(s):  
Canine Distemper Virus ◽  
Duplex Pcr ◽  
Canine Distemper ◽  
Pcr Assay ◽  
Canine Coronavirus ◽  
One Step ◽  
Duplex Pcr Assay

scholarly journals One-Step Rapid Quantification of Serum Neutralizing Antibody after COVID-19 Vaccination by a High-Throughput Nanoplasmonic Sensor Platform

2021 ◽  
Author(s):  
Liping Huang ◽  
Ying Li ◽  
Luo Changyou ◽  
Nadia Touil ◽  
Hicham el Annaz ◽  
...  
Keyword(s):  
High Throughput ◽  
Vaccine Development ◽  
Limit Of Detection ◽  
Neutralizing Antibody ◽  
Serum Samples ◽  
Igg Antibody ◽  
Plasma Therapy ◽  
One Step ◽  
The One ◽  
Rapid Quantification

ABSTRACTThe COVID-19 vaccination efficacy depends on serum production level of the neutralizing IgG antibody (NA) specific to the receptor binding domain of SARS-Cov-2 spike protein. Therefore, a high-throughput rapid assay to measure the total SARS-CoV-2 NA level is urgently needed for COVID-19 serodiagnosis, convalescent plasma therapy, vaccine development, and assessment. Here, we developed a nanoplasmonic immunosorbent assay (NanoPISA) platform for one-step rapid quantification of SARS-CoV-2 NAs in clinical serum samples for high-throughput evaluation of COVID-19 vaccine effectiveness. The NanoPISA platform enhanced by the use of nanoporous hollow gold nanoparticle coupling was able to detect SARS-CoV-2 NAs with a limit of detection of 0.1 ng/mL within 15 min. The one-step NanoPISA for SARS-CoV-2 NA detection in clinical specimens yielded good results, comparable to those obtained in the gold standard seroneutralization test and the surrogate virus neutralizing ELISA. Collectively, our findings indicate that the one-step NanoPISA may offer a rapid and high-throughput NA quantification platform for evaluating the effectiveness of COVID-19 vaccines.

A Ratiometric Fluorescent Probe Based on Carbon Dots and Bimetallic Nanoclusters for the Assay of Copper Gluconate and Copper Sulfate

NANO ◽  
2021 ◽  
pp. 2150114
Author(s):  
Yucong Fan ◽  
Weihua Yu ◽  
Yue Hu ◽  
Yunwen Liao ◽  
Xiaohui Jiang ◽  
...  
Keyword(s):  
Fluorescent Probe ◽  
Carbon Dots ◽  
Limit Of Detection ◽  
Molar Ratio ◽  
Cupric Sulfate ◽  
Selective Determination ◽  
Bimetallic Nanoclusters ◽  
One Step ◽  
The One

Doping Ag-enhanced and glutathione-stabilized Au nanoclusters (GSH–Ag/AuNCs) were prepared by the one-step ultraviolet radiation combined with microwave heating method. The effects of the molar ratio of Au–Ag and different types of energy suppliers on the fluorescent performance of GSH–Ag/AuNCs were studied in detail. After that, a new ratio fluorescent probe (RF-probe) based on the mixing of GSH–Ag/AuNCs with carbon dots (CDs) was designed for sensitive and selective determination of copper gluconate (CG) and cupric sulfate (CS). For the CDs–GSH–Ag/AuNCs RF-probe, the fluorescence (FL) of CDs (at 440[Formula: see text]nm) and that of alloy nanoclusters (NCs) (at 605[Formula: see text]nm) were, respectively, unaffected and strongly quenched in the presence of CG/CS at [Formula: see text][Formula: see text]nm coming from the dynamic quenching process. Corresponding linear ranges and limit of detection (LOD) of the RF-probe for the CG/CS assay were estimated to be 0.17–6.20/0.17–5.62[Formula: see text][Formula: see text]mol/L and 16.80/15.95[Formula: see text]nmol/L, respectively. Furthermore, the proposed RF-probe was successfully used for the assays of CG in CG tablets and CG additive, and CS in infant formula and CS additive, respectively.

scholarly journals One-step triplex PCR/RT-PCR to detect canine distemper virus, canine parvovirus and canine kobuvirus

2019 ◽  
Vol 81 (7) ◽  
pp. 1040-1042 ◽  
Author(s):  
Dafei LIU ◽  
Fei LIU ◽  
Dongchun GUO ◽  
Xiaoliang HU ◽  
Zhijie LI ◽  
...  
Keyword(s):  
Canine Distemper Virus ◽  
Canine Parvovirus ◽  
Canine Distemper ◽  
Rt Pcr ◽  
One Step ◽  
Canine Kobuvirus

scholarly journals Detection of canine distemper virus (CDV) through one step RT-PCR combined with nested PCR

2001 ◽  
Vol 2 (1) ◽  
pp. 59 ◽  
Author(s):  
Yong Hwan Kim ◽  
Kyu Woan Cho ◽  
Hwa Young Youn ◽  
Han Sang Yoo ◽  
Hong Ryul Han
Keyword(s):  
Nested Pcr ◽  
Canine Distemper Virus ◽  
Canine Distemper ◽  
Rt Pcr ◽  
One Step

scholarly journals Development of recombinase polymerase amplification assays for rapid and visual detection of canine distemper virus infecting giant panda

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Pei Huang ◽  
Yue Yu ◽  
Xianyong Meng ◽  
Tiecheng Wang ◽  
Feihu Yan ◽  
...  
Keyword(s):  
Canine Distemper Virus ◽  
Giant Panda ◽  
Limit Of Detection ◽  
Natural Infection ◽  
Rapid Test ◽  
Recombinase Polymerase Amplification ◽  
N Gene ◽  
Clinical Samples ◽  
Canine Distemper ◽  
Giant Pandas

Abstract Background Canine distemper virus (CDV) is an enveloped negative-strand RNA virus that exhibits a high mutation rate and continuously expands the range of hosts. Notably, CDV has infected giant panda with spill over from viral reservoirs in canines. Giant pandas (Ailuropoda melanoleuca), especially captive pandas, are known to be susceptible to natural infection with CDV. The high fatality rate of CDV poses a serious threat to the safety of the giant panda population. However, vaccines or drugs for canine distemper in giant pandas have not been developed to date. Therefore, a rapid test that can achieve accurate onsite detection of CDV is important to enable the timely implementation of control measures. In this study, we established a nucleic acid visualization assay for targeting the CDV N gene by using combines reverse transcription recombinase polymerase amplification with a closed vertical flow visualization strip (RT-RPA-VF). Results The RT-RPA-VF assay does not require sophisticated equipment, and it was determined to provide rapid detection at 35 °C for 30 min, while the limit of detection was 5 × 101 copies/μl RNA transcripts and 100.5 TCID50 ml− 1 viruses. The results showed that the assay was high specific to CDV and had no cross-reactivity with other viruses infecting the giant panda. Compared with RT-qPCR, RT-RPA-VF assay had a sensitivity of 100% and a specificity of 100% in 29 clinical samples. The coincidence rate between RT-RPA-VF and RT-qPCR was 100% (kappa = 1), indicating that the RT-RPA-VF assay possessed good diagnostic performance on clinical samples. Conclusions The RT-RPA-VF provides a novel alternative for the simple, sensitive, and specific identification of CDV and showed great potential for point of care diagnostics for captive and wild giant panda.

Sign in / Sign up

Export Citation Format

Share Document