Name: titration of human coronaviruses using an immunoperoxidase assay
Uploaded: 2008-04-28T22:00:00
Duration: 12 min
Description: Watch this Scientific Journal Video about Titration of Human Coronaviruses Using an Immunoperoxidase Assay at JoVE.com

이 실험 방법에 대한 전체 텍스트 프로토콜에서 사용할 수 있습니다 <a href="http://www.springerprotocols.com/">스프링거 프로토콜</a> .

The authors have nothing to disclose.

A correction was made to: <a href="http://www.jove.com/video/751/">Titration of Human Coronaviruses Using an Immunoperoxidase Assay</a>. A revised abstract was republished due to a publisher error.
Revised Abstract:
Determination of infectious viral titers is a basic and essential experimental approach for virologists. Classical plaque assays cannot be used for viruses that do not cause significant cytopathic effects, which is the case for prototype strains 229E and OC43 of human coronavirus (HCoV). Therefore, an alternative indirect immunoperoxidase assay (IPA) was developed for the detection and titration of these viruses and is described herein. Susceptible cells are inoculated with serial logarithmic dilutions of virus-containing samples in a 96-well plate format. After viral growth, viral detection by IPA yields the infectious virus titer, expressed as 'Tissue Culture Infectious Dose 50 percent' (TCID50). This represents the dilution of a virus-containing sample at which half of a series of laboratory wells contain infectious replicating virus. This technique provides a reliable method for the titration of HCoV-229E and HCoV-OC43 in biological samples such as cells, tissues and fluids. This article is based on work first reported in Methods in Molecular Biology (2008) volume 454, pages 93-102.
Original Abstract:
Calculation of infectious viral titers represents a basic and essential experimental approach for virologists. Classical plaque assays cannot be used for viruses that do not cause significant cytopathic effects, which is the case for strains 229E and OC43 of human coronavirus (HCoV). An alternative indirect immunoperoxidase assay (IPA) is herein described for the detection and titration of these viruses. Susceptible cells are inoculated with serial logarithmic dilutions of samples in a 96-well plate. After viral growth, viral detection by IPA yields the infectious virus titer, expressed as "Tissue Culture Infectious Dose" (TCID50). This represents the dilution of a virus-containing sample at which half of a series of laboratory wells contain replicating virus. This technique is a reliable method for the titration of HCoV in biological samples (cells, tissues or fluids).

A correction was made to: <a href="http://www.jove.com/video/751/">Titration of Human Coronaviruses Using an Immunoperoxidase Assay</a>. A revised abstract was republished due to a publisher error.

Erratum: Titration of Human Coronaviruses Using an Immunoperoxidase Assay

titration of human coronaviruses using an immunoperoxidase assay

전염성 바이러스 titers의 계산 virologists위한 기본적인 필수 실험 방법을 나타냅니다. 고전 플라크의 assays는 종자 229E 인간 coronavirus (HCoV)의 OC43의 경우는 상당한 cytopathic 효과를 발생하지 사용할 수있는 최신 바이러스 예방 프로그램으로 사용할 수 없습니다. 다른 간접 immunoperoxidase 분석 (IPA)는 본 이들 바이러스의 감지 및 적정에 대해 설명되어 있습니다. 민감한 세포는 96 - 웰 플레이트에 샘플 시리얼 로그 dilutions와 주사를하고 있습니다. 바이러스성 성장 후, I​​PA의 바이러스 검출은 &quot;조직 문화 전염성 선량&quot;(TCID50)로 표현 전염성 바이러스 titer를 산출. 이것은 실험실 우물 일련의 절반이 복제 바이러스를 포함하는에서 바이러스가 포함된 시료의 희석을 나타냅니다. 이 기술은 생물 학적 샘플에서 HCoV의 적정 (세포, 조직 또는 체액)에 대한 안정적인 방법입니다.

Watch this Scientific Journal Video about Titration of Human Coronaviruses Using an Immunoperoxidase Assay at JoVE.com

Titration of Human Coronaviruses Using an Immunoperoxidase Assay

이 비디오에서 우리는 immunoperoxidase 분석으로 알려진 효소 항원 검출 기법을 사용하여 바이러스 감지 및 titering위한 대체 방법을 보여줍니다. 여기, 우리는 여러분의 바이러스 샘플을 수집하는 방법을 보여 테스트 세포를 준비하고, 바이러스 titer를 결정하는 일련 dilutions를 사용하여 최종 immunoperoxidase 분석됩니다.

Immunoperoxidase 분석을 사용하여 인간 Coronaviruses의 적정

Determination of infectious viral titers is a basic and essential experimental approach for virologists. Classical plaque assays cannot be used for ...

Research

JoVE Journal

Biology

Homemade Site Directed Mutagenesis of Whole Plasmids

전체 Plasmids의 수제 사이트 감독 돌연변이 유발

Site directed mutagenesis of whole plasmids is a simple way to create slightly different variations of an original plasmid. With this method the cloned ...

연구

생물학

An Allele-specific Gene Expression Assay to Test the Functional Basis of Genetic Associations

유전자 협회의 기능 기초를 테스트하는 Allele 특정 유전자 발현 분석

The number of significant genetic associations with common complex traits is constantly increasing. However, most of these associations have not been ...

Imaging C. elegans Embryos using an Epifluorescent Microscope and Open Source Software

Imaging C. elegans Embryos using an Epifluorescent Microscope and Open Source Software

이미징 C. 엘레간스</emEpifluorescent 현미경과 오픈 소스 소프트웨어를 사용> 태아

Cellular processes, such as chromosome assembly, segregation and cytokinesis,are inherently dynamic. Time-lapse imaging of living cells, using ...

Analysis of SNARE-mediated Membrane Fusion Using an Enzymatic Cell Fusion Assay

효소 세포 융합 분석을 사용하여 올가미로 인한 막 퓨전 분석

The interactions of SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor)  proteins on vesicles (v-SNAREs) and on target membranes ...

Biochemical Titration of Glycogen In vitro

Biochemical Titration of Glycogen In vitro

글리코겐의 생화학 적정<em&gt; 체외</em

Glycogen is the main energetic polymer of glucose in vertebrate animals and plays a crucial role in whole body metabolism as well as in cellular ...

Measuring the Effects of Bacteria on C. Elegans Behavior Using an Egg Retention Assay

Measuring the Effects of Bacteria on C. Elegans Behavior Using an Egg Retention Assay

에 박테리아의 효과를 측정<em&gt; C. 엘레</em계란 보존 분석을 사용하여&gt; 행동

C. elegans egg-laying behavior is  affected by environmental cues such as osmolarity1 and  vibration2. In the total absence of food C. elegans also cease ...

High-throughput Titration of Luciferase-expressing Recombinant Viruses

루시 페라 제 발현 재조합 바이러스의 높은 처리량의 적정

Standard plaque assays to determine infectious viral titers can be time consuming, are not amenable to a high volume of samples, and cannot be done with ...

A Method for Selecting Structure-switching Aptamers Applied to a Colorimetric Gold Nanoparticle Assay

색채 금 나노 입자 분석에 적용 구조 전환 앱 타머를 선택하는 방법

Small molecules provide rich targets for biosensing applications due to their physiological implications as biomarkers of various aspects of human health ...

In Vivo Study of Human Endothelial-Pericyte Interaction Using the Matrix Gel Plug Assay in Mouse

In Vivo Study of Human Endothelial-Pericyte Interaction Using the Matrix Gel Plug Assay in Mouse

마우스의 매트릭스 젤 플러그 분석을 사용하여 인간의 내피 - 주연 세포 상호 작용의 생체 내 연구에

Angiogenesis is the process by which new blood vessels are formed from existing vessels. New vessel growth requires coordinated endothelial cell ...

LDL Cholesterol Uptake Assay Using Live Cell Imaging Analysis with Cell Health Monitoring

LDL 콜레스테롤 이해 분석 결과 라이브 셀 이미징 분석 셀을 사용 하 여 상태 모니터링

The regulation of LDL cholesterol uptake through LDLR-mediated endocytosis is an important area of study in various major pathologies including metabolic ...