Detection of Virus

DETECTION OF VIRUSES - PHYSICAL, BEROLOGICAL AND BIOLOGICAL OF MALICULAR METHADS Detection Of Virus: It is defined at the action of detecting presencelabsence of vious in the given sovice car) dample The ability to determine the presence of love can be dow by 3 methods, they are T. Physical Method 2. Serological method 3. Bialogical Imoleculas Method 1. PHYSICAL METHODS:- It refers to detection of virus by Using Instruments (x) equipments such as, a. Light Microscope b. Electron Microscope f. x-ray Crystallography d, centrifugation a Light Microscope: The siges of thoat visions are resolution of light microscopes. beyond the limits of →But, light microscopy hac useful applications in detecting virus-infected cells. for Example, by observing cytopathic effects (bo) by deiecting a fluorescent dye linked to antibody matkules thai have bound to a virus antigen [fluorescence Microscopy). 6. Electron Microscope: Analysis of virus (detection of vieve in a given Sample I specimen can be clone by using election microscope. →In this meltiod, beam of electrons from election gun are used for detect Virus Lange magnification are achievable with a transmiste electron microscope, but ultra thin section of Viruses are prepared The Sectioning is done with diamond of glase Knife and embedded (fring) Now, Stained with heavy metal containing compounde [potassium phosphotungstate and Aminanium molybdate] to make certain part dense and inserted in vaccum Chamber of microstepe →Then by paccing 100,000 volt of electron beam on ite Section we can visualize and magnity ultra section of viove particle. F. X-ray Crystallography: →x-ray Crystallography is another technique that detect the nous by reveating detailed information about three dimensional Structure of virions [DNA, proteins 4 eNA-prottin complerts). →This technique requies the production of a crystal of the virion/molecule under study. →The crystal is placed in a beam of x-rays which are diffracted by repeating arrangements of aloms in the Crystal so by analyu's of diffraction pattern allows the relative poulbion of Vision/virus/malecult to be determined. Contrifugation wowee can be detected by using centrifugation process The Virut in a mard population con) rival particles in water (liquids can be detected (asseparated by centringatian contol fugation tavolves alternating cycles of low-speed Emisingation, after which most of virus is stilu in the Supernatant, and in high-speed centrifugation, the virus It in pellet form 2 SEROLOGICAL METHOD This teste can be used to detect viral and bacterial antigens and antibodies to help diagnose diseases Rock Check Immune Status. →Serological methods Includes, A. ELIGA 4. Immunofluorescence & Haemagglutination inhibition the Radio immunoassay 4. Camptement (zation test P. ELISA Enzyme tinked immuno sorbent Alsay- It is used to derect the Vival ontibodies in blood Sample In ELISA Kit [micreditse well) se coared with ag and they are very specific to adhere to particular type of thb. To detect the wawa in srium, the shown in to heatfer lott microlitre well If virus ab's present in Strom. If coll dbers to Ag Labelled antchody is added to detect web] It is labelled with enzyme cwhich can react is th Sultan Substrate to produce Vilible change for ago. Ag Coated plate + patient serven add enzyme abriled Ab incubute and then add tubstrate in Bra Badie Immune Assay →eit is similar to crise, but antibody is tagged cáth an Hetople out aften 125 radialsofape It will detect the viral Ag Content in a Jample Haemagglutination inhléilion test Harmegglutination refers to compling of red blood cells. →But if a Star Contains του → that Viral Aq can be detected by inhibition of Harmogglutination. d compterpent fixation test: →tils ad protein (made by sharp erythrocytes le fo →Generally, in corvmplement fixation test, Ab attached with complement will form Ag-Ab iniriachan in blood Plyos of Rec But in detection method, this OFT test ward to delect the Visal Ay via complement fixation ie, if sampe Contains viral ng aftes adding the complement is fixed and red blood cells do not haemolysed 16. Neutralizations Neutralization is other serological method to denct the Viral Ag in a Serum by certain antibodies cated neutralizing antibodice. This Abis seact with visal Ag and neutralize them There are a types of Nutsatization. 1. Toxic Neutralization 8. Visal Neutralization Tone Neutralization refers to antibody produced in response to bacterial toxold and that Ab neutralize to toxin. vial Neutralization Vrat mutralization is the process dusing which specifle Ab blad to the wave partical and inactivate them. 3. MOLECULAR BIOLOGY METHODS: It is the branch of biology that deals with the Simitest and function of the macro-moleculee. kg: protein and nuckle aeld →This technique aled to detect the one/Hows Componente [DNATRNA Protein] By using molrculas method wry small amount) pation of pathogen can be detection Matniy by PCR & blotting techniques and Gel Mectrophorete PCR: Polymerase Chain reaction (PER)Ts a technique used in molecular blology to amplify a single copy on few capire of a segment of Due across several orden of magnitude, generating theutards to millions of copies of a particular DNA Ingoence It to developed by kary Mulls in the year 1965. Gelectrophoras: It is a laboratory method wied to ceparate mistuse of Due, we (1) proteins according to molecules F. Blotting technique:- It is a method of transferring proteins, DANA (D) ANA onto a Carsler. [for £q: a nitracellulose (or) nylon membrane) →Blotting techniques are of different types Southern blotting wasten blotting Misathan blotting Southern blatting: Laboratory method used to detect specifle DNA molecules from among a many other DNA moleculee 8. Western blotting: It is used to detect specific proteins in a given cample Northern blotting: It is used to detect the Run from givere Sample process to detect virus i genetic maternal: Sample CONA) POR ↓ Gel Electrophoresis Transfer to Nylon membrane for blotting fidelitian of probe Detection Sample (ENA) RT-PCR Gel Electrophoreshs Transfer to Nylon membrane for bletting addition of probe Lerechan