<< Chapter < Page Chapter >> Page >

When viruses infect cells, they often cause damage ( cytopathic effects ) that may include lysis of the host cells. Cytopathic effects can be visualized by growing host cells in a petri dish, covering the cells with a thin layer of agar, and then adding virus (see Isolation, Culture, and Identification of Viruses ). The virus will diffuse very slowly through the agar. A virus will enter a host cell, proliferate (causing cell damage), be released from the dead host cell, and then move to neighboring cells. As more and more cells die, plaques of dead cells will form ( [link] ).

During the course of a viral infection, the patient will mount an antibody response to the virus, and we can quantify those antibodies using a plaque reduction assay . To perform the assay, a serial dilution is carried out on a serum sample. Each dilution is then mixed with a standardized amount of the suspect virus. Any virus-specific antibodies in the serum will neutralize some of the virus. The suspensions are then added to host cells in culture to allow any nonneutralized virus to infect the cells and form plaques after several days. The titer is defined as the reciprocal of the highest dilution showing a 50% reduction in plaques. Titer is always expressed as a whole number. For example, if a 1/64 dilution was the highest dilution to show 50% plaque reduction, then the titer is 64.

The presence of antibodies in the patient’s serum does not tell us whether the patient is currently infected or was infected in the past. Current infections can be identified by waiting two weeks and testing another serum sample. A four-fold increase in neutralizing titer in this second sample indicates a new infection.

A photograph of wells showing a smooth purple background with white spots.
In a neutralization assay, antibodies in patient serum neutralize viruses added to the wells, preventing the formation of plaques. In the assay pictured, the wells with numerous plaques (white patches) contain a low concentration of antibodies. The wells with relatively few plaques have a high concentration of antibodies. (credit: modification of work by Centers for Disease Control and Prevention)
  • In a neutralization assay, if a patient’s serum has high numbers of antiviral antibodies, would you expect to see more or fewer plaques?

Immunoelectrophoresis

When a patient has elevated protein levels in the blood or is losing protein in the urine, a clinician will often order a polyacrylamide gel electrophoresis ( PAGE ) assay (see Visualizing and Characterizing DNA, RNA, and Protein ). This assay compares the relative abundance of the various types of serum proteins. Abnormal protein electrophoresis patterns can be further studied using immunoelectrophoresis (IEP) . The IEP begins by running a PAGE. Antisera against selected serum proteins are added to troughs running parallel to the electrophoresis track, forming precipitin arcs similar to those seen in an Ouchterlony assay ( [link] ). This allows the identification of abnormal immunoglobulin proteins in the sample.

IEP is particularly useful in the diagnosis of multiple myeloma , a cancer of antibody-secreting cells. Patients with multiple myeloma cannot produce healthy antibodies; instead they produce abnormal antibodies that are monoclonal proteins (M proteins). Thus, patients with multiple myeloma will present with elevated serum protein levels that show a distinct band in the gamma globulin region of a protein electrophoresis gel and a sharp spike (in M protein) on the densitometer scan rather than the normal broad smear ( [link] ). When antibodies against the various types of antibody heavy and light chains are used to form precipitin arcs, the M protein will cause distinctly skewed arcs against one class of heavy chain and one class of light chain as seen in [link] .

Questions & Answers

write the life cycle of HIV
Firomsa Reply
describe the internal and external structure of prokaryotic cell in terms of there appearance and functions
Lenia Reply
compare and contrast similar structures found in prokaryotic and eukaryotic cells
Lenia
control of microorganisms
ISTEFAZUL Reply
how can a doctor treat a person affected by endospore forming bacteria in his/her wound?
Mambo Reply
a 28 years old woman come to your clinic with complain of fever painful genital blisters which express clear fluid when ruptured burning sensation around the bristers what is the diagnosis?
Ramadhani Reply
genital herpes caused by a virus called herpe simplex virus
Doris
what is the reference between selective medium and differential medium?
Tony Reply
the micro flora of air is transient why
Princess
Hello.... Am new here
essien Reply
welcome essien
Muhammad
welcome essien
Beka
classification of gram positive
lissa Reply
classify gram positive
lissa
catalase test is done to differentiate between staph and strep. as staph is catalase positive while strep is catalase negative then staph is differentiate further on coagulase positive and coagulase negative. staph aureus is coagulase positive while staph epidermidis is coagulase negative.
Muhammad
gram positive staph is further differentiated on sensitivity test and manitol salt fermentation test while gram positive strep is differentiated on hemolysis pattern
Muhammad
more about gramm positive
lissa
please
lissa
ok
Blessing
am back any gist
Blessing
taxonomy
Rahul
can i use the graph of bacterial growth for my master's thesis?
Christoph Reply
facultative anaerobic bavteria gives uniform turdibity in nutrient broth why?
Princess Reply
oils and waxes are not sterilized in autoclave
Princess
what is the function of paraffin
Muhammad
@ MAHI because they can grow all over the media from surface to the bottom as they can utilize oxygen or conduct fermentation in its absence.
Online
2.@MAHI Autoclave uses steam under pressure. The steam cannot penetrate through oil and wax. Thus, dry heat sterilization is preferred than autoclave .
Online
thnku
Nadiya
how nutirent agar can converted into blood agar
Princess
thank you
Princess
take a nutrient agar ........and add 5ml blood and put in it ...........!
Nadiya
nutrient agar +blood 5ml+ distilled water =blood agar .
Nadiya
mixed wellllll
Nadiya
hi
Rahul
why agar is not a neutrient source?
Shimul
because it's a general purpose agar only use for growing bacteria
Richa
way is antigen
opaleye
what is antigen
opaleye
antigen is a foreign body that cause activation of antibody?
ASNAKE
Antigen is a toxin or other foreign substance which induces an immune response in the body, especially the production of antibodies.
Shimul
thanks for that
opaleye
what is pharmacology
Luyenu
a science that studies about the drug
ASNAKE
hello ever one
Sayid
y
Abdul
Work hard
Anigor
y
Abdul
thanks alot
Luyenu
What is the major different between gram positive and gram negative bacteria.
Asikur
bacteria kya hai
Mala
content of their cell wall
Doris
koi mujhse basic microbiology ki study k Lea book suggest kro .
Richa
Harley prescott
Vikas
what are the five predisposing factors of opportunistic infections
Asher Reply
1. long term exposure to diseases, that the body immunity weakens.
prosper
2. Nature Of some diseases to weaken body immunity such as, AIDS.
prosper
3. Use of immuno-suppresive drugs.
prosper
anyone... need a hand here 🙋😞
prosper
Low immune is the major cause eg for kaposis sarcoma
Davismith
suitable conditions e.g temp,
Muhammad
what os gnormal flora
kifayat Reply
morphological and genetic classification of bacteria
Potter Reply
what is the morphogical
Bupe
h
Sudhir
morphologycal is defined as relating to the branch of biology that deals with the form of living organisms,and with relationship between their structures.
Nadiya
relating to the form or structure of things .
Nadiya
what is the meaning of lessions
oluwa
i am frim Afghanistan.
wahidullah Reply
iam from iraq
Haidar
and i'm from pakistan
Muhammad
Nice
Amina
what's about the qualification
Muhammad
Hlo
Asiya
Hmm
Number
Almustafa Muhammad from Nigeria
Mujtafa
am ustaz Abdulsalam from Ghana
ustaz
am Bupe Chifita from Zambia
Bupe
what is microbiology
Bupe
is the study of invisible micro organism
Mujtafa
microbiology is the study of very too small organisms which we can't see with naked eyes ......in other words ....study of microorganism is called microbiology.
Nadiya
Hello! How Can Microorganisms Be Isolated From The Skin?.. 5 ways if possible. (briefly)
prosper
from skin swabbing is the prefer method worldwide
Muhammad
thanks Muhammad Nauman very much!
prosper
hi
Meek
hi Meek
prosper
are there any other ways pals?
prosper
hi prosper
Meek
hello Meek Mild.
prosper
Any question or ideas, Meek?😊
prosper
and if a patient have any blister or abscess on the skin then first cut the blister or abscess. inside material is taking by syringe.
Muhammad
in case of fingernail or toenails simply cut a small piece of nail and culture on the appropriate media at a required temperature and time
Muhammad
Again, thanks Mr. Muhammad! I appreciate the help.
prosper
you welcom prosper 😊
Muhammad
what are the media use for skin swab?
Sieh-Mlanwin
blood and macckonkey mostly usededia for skin bacteria
Muhammad
used media*
Muhammad
@ SIEH mostly fungal infection occurs in skin so SDA ( Sabouraud Dextrose Agar) or MEA ( Malt extract agar) are common.
Online
thanks
Sieh-Mlanwin
Microbiologicam source of vitamins
Bachi Reply
what do you mean?
Md
that's example?
Md
hi everyone
Muhammad
such as. c/s media
Md
hmm hlw
Md
just tell me about that ?
Md
where are u from
Muhammad

Get the best Microbiology course in your pocket!





Source:  OpenStax, Microbiology. OpenStax CNX. Nov 01, 2016 Download for free at http://cnx.org/content/col12087/1.4
Google Play and the Google Play logo are trademarks of Google Inc.

Notification Switch

Would you like to follow the 'Microbiology' conversation and receive update notifications?

Ask