CMV diagnosis

Tim at hgmp.mrc.ac.uk Tim at hgmp.mrc.ac.uk
Tue Mar 14 09:43:36 EST 2000


Historically and typically, by observing distinct cytopathic effect
(CPE) in
human fibroblast cells in tissue culture tubes.  This is slow and may
take up
to 4 weeks to display the CPE.  Our hospital also utilizes a "shell
vial" and
FITC-conjugated monoclonal antibody staining procedure. In this
procedure the
patient's specimen is added to a shell vial that has a round coverslip
at the
bottom of the vial.  The coverslip is layered with human fibroblasts.
The
patient's specimen is added to a set of shell vials and the shell vials
are
centrifuged for about 1 hour.  Culture media is added to the shell vial
and
the shell vial is incubated at 37 degrees.  At different time points
over the
course of 1-7 days post inoculation, a shell vial is removed from the
incubator and stained with a FITC-conjugated monoclonal antibody
developed
against a CMV early nuclear antigen.  The stained round coverslips are
mounted
on a slide and perused under a UV scope for cells that show distinct
positive
nuclear staining (almost like "glowing kidney-shaped" nuclei).
Specimens that
we have used for both of these procedures include human white blood
cells
harvested from a buffy coat separation, urine, and inguinal swabs
immersed in
viral transport medium.
A "positive" IgM antibody test run on serum indicates an active or
recent
infection.

I've been a few years away from this, but I'm told that some kits have
been
developed that utilize PCR to detect CMV (these are Roche kits).  I
suspect
that other companies have probably developed various configurations of
direct
and indirect fluorescent microscopy for the detection of CMV antigen.

daemon at hgmp.mrc.ac.uk, [mailto:daemon at hgmp.mrc.ac.uk], On, Behalf, Of
wrote:

> How is CMV diagnosis usually done? Specifically, which lab tests can
be
> performed?
>
> Thanks!
>
> Paula Lavery
> Transplant Immunology Lab.
> Royal Victoria Hospital
> McGill University
> Montreal (QC) Canada
>
> "If we knew what we were doing it wouldn't be called research, would
> it?"
> -- Albert Einstein
>
> ---





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