Wednesday, February 26, 2014

Cell Culture Sensitivity for Various Viruses

Part of what I want to do with this blog is put up some of our protocols and equipment that we use. The first protocol discussion will be on our virus culture algorithm. This is a guideline that we use in the lab to determine what potential virus we are dealing with.

“If you need to convince yourself that it is positive, it’s negative”
 
Key
RMK - Rhesus Monkey Kidney
HEp-2 - HeLa derivitive
HFS - Human Foreskin
MRC-5 - Human Fetal Lung Fibroblast
BGMK - Buffalo Green Monkey Kidney
Caco-2 - Human Epithelial Colorectal Adenocarcinoma
MDCK - Madin-Darby Canine Kidney
RD - Rhabdomyosarcoma

Amount of Cytopathic Effect (CPE)
0 - None
+ - A little
++ - More
+++ - Most 
 
Cell Type
Virus
RMK
HEP-2
HFS
MRC-5
BGMK
Caco-2
MDCK
RD
 
Herpes Simplex Virus -1
+
+
+++
+++
++
+
+
++
On HFS, HSV-1 appears as smaller rounded cells, very few or no vacuoles, cells appear to be branching out
Herpes Simplex Virus -2
+
+
+++
+++
++
+
+
++
On HFS, HSV-2 appears as larger rounded cells; many cells have multiple large vacuoles
Varicella Zoster Virus
+
0
+++
++
0
0
0
0
Generally remains focal, without spreading to entire monolayer.  “Lacy” appearance of CPE
Cytomegalo
virus
0
0
++
++
0
0
0
0
Very slow growing, limited foci of large round cells
Adenovirus
++
++
+
+
0
++
0
0
Large rounded cells similar to HSV
Influenza A
++
0
0
0
0
++
++
0
Depending on the strain each year it may grow best on RMK, MDCK, or Caco-2 cell line.  Some years it may not grow on one of those cell lines at all. Can do hemadsorption with guinea pig RBC’s
Influenza B
++
0
0
0
0
++
+++
0
Similar to influenza A; often shows up late winter, after Flu A has peaked. Can do hemadsorption with guinea pig RBC’s
Respiratory Syncytial Virus
+
+++
+
0
0
+
0
0
Syncytia look like “drops of oil”
Coxsackie A
+
0
0
0
0
+
0
+++
Smaller rounded up cells.  May grow only on RD or Caco-2
Coxsackie B
+++
++
0
0
+++
++
0
0
Smaller rounded up cells. 
Echovirus
+
0
+++
+++
?
+++
0
+
Smaller rounded up cells.  Some strains (echo-11) grow well on BGMK
Parainfluenza
+++
0
0
0
0
0
0
0
Often does not show CPE despite virus growing.  Can do hemadsorption with guinea pig RBC’s
Rhinovirus
+
0
++
++
0
0
0
0
Spread out CPE of various sized, highly refractive, rounded cells
Mumps
+++
0
0
0
?
?
0
0
From buccal swabs or urine.  Confirm with PCR
Reovirus
++
0
0
0
++
0
0
0
Common environmental isolate.  BGMK cells appear “grainy” with a perinuclear ring.
 
I also want to encourage readers if they have questions, to use the comments section!

Lance D. Presser has a PhD in Microbiology and Immunology and currently is a Public Health Laboratorian.

Hire Lance as a consultant using Zintro.
 



1 comment:

  1. Nice job. Any information on Morbilliviruses and arboviruses?

    ReplyDelete