Adenovirus Articles


Adeno Virus Testing at
the University of Illinois

The diagnostic Electron Microscopy Services at the University of
Illinois (http://www.uiuc.edu) are located at the College of Veterinary
Medicine (http://www.cvm.uiuc.edu), Center for Microscopic Imaging(CMI)
(http://treefrog.cvm.uiuc.edu)

The test performed at CMI is that of visual recognition of the virus
itself, a test technique commonly known as “negative staining.” In this
test, the sample is mixed with a specialized fixative solution, spun
down by different centrifugation techniques, applied to a specialized
mesh covered with a plastic film, and then stained with a heavy metal
stain that flows around the virus, sometimes into the virus and allows
the virus to be seen in relief against a dark background. Characteristic
markings of a virus can be identified with this technique by observation
under an electron microscope.


Adeno virus can be reported by several methods. In addition to negative
staining, the actual visual examination of a sample with an electron
microscope, there is a PCR test that looks for the DNA or RNA of an
organism. The PCR is the more sensitive and often the more expensive
test and holds more possibilities for false positives. The negative
stain test is less sensitive and more prone to false negatives.

Many people do not realize that false positives, false negatives, and
sensitivity are factors involved in all laboratory testing methods.
Often in laboratory testing the less expensive broad spectrum test is
done first, and the more specific test is used for confirmation, or in
this case to test the negative test results of the screening test.

One advantage of the visual method is that other viral agents can be
seen and reported.
The advantage of PCR is that it is very specific.


How to Test:
* Obtain a fixation kit. Email Lou Ann Miller at CMI
(lamiller@uiuc.edu) to request the number of vials necessary for
your needs. Then send a check for $10 made out to the University of
Illinois. This $10 fee is for the fixative kit ONLY, the actual testing
will be $23.45/ sample and is paid with a check at the time of sending
the sample. One or more than one fixative vial can be requested for the
same fee. The fixative should be kept in the refrigerator upon arrival
and should not be allowed to touch skin; use gloves provided when
handling the vials. Instructions for collection are included in the kit.


Send kit request to:
Lou Ann Miller, Center for Microscopic Imaging
College of Veterinary Medicine
Room 1212 VMBSB
2001 S Lincoln Ave
Urbana, IL 61802
217-244-1567
lamiller@uiuc.edu (Email is preferred for correspondence)

Collection
1. The fecal sample is collected, and must be fresh, less than 20
minutes old. The sample can be immediately frozen in a plastic sealed
bag and kept while others are collected, or the sample can be placed
immediately into fixative and stored for only a few days at refrigerator
temperatures before sending out. Frozen samples are to be placed into
the fixative to thaw prior to sending.

2. The sample is to be sent into the lab well sealed in several layers
of Ziplock bags and with an ice pack. Use FedEx or UPS; 2-day delivery
is fine. Do not send by mail because the sample will not be as speedily
delivered.

3. Other checks:
* Is the fixative clear, and was it kept out of light in the refrigerator?

* Is the name, identifying number etc. marked clearly on the collection
tube with permanent ink?

* Is the sample double sealed with zip bags?

* If you are certifying the testing procedure with Pogana Testing
services, are the proper forms and paperwork included?

* Is the client information (name, email, address, phone) included on
the inside of the box?

* Is the paraffin provided wrapped around the top of the tube?

* Is the check for the test procedure ($23.45/ sample) made out to the
“University of Illinois” included in the shipment?

* Are the tubes on an ice pack?  Do not use dry ice.

* Please check via email that the package you sent has arrived.


About the Testing
4. The negative stain procedure is done on Mondays, Wednesdays, and
Fridays, starting first thing in the morning and requires about a half
day’s worth of centrifugation procedures. Usually the test is done with
in 2 days of receiving a sample.

5. Reporting. CMI will send you the results via email, and if desired a
hard copy in the mail. If you are certifying the results, and give
permission, the results will also be sent to Pogana Testing. Any virus
seen will be reported, and if there is indication of possible parasites
that need testing for, this information will also be given to you.
However, this test is not prepared to be specific or reliable for parasites.

In this type of visual testing, a positive is always reported out as
“adeno like virus particles seen.” This is standard for all viral
procedures in this lab. A negative will be reported as “no viral
particles seen.” With a positive, a semi quantitative modifier will also
be added: rare, occasional, moderate, many. This testing is visual, and
this phrasing is used in reporting.


For large batches of testing:
Some clutches can have pooled samples, two to three samples pooled
together and tested. This does save money, and is done with herds, if
the pool is positive. However, one does not know which or if all of the
samples is positive.

The Adeno Virus
Many questions about adeno virus are better directed to a virologist or
epidemiologist.

It is a hardy virus, it can live on dry surfaces, get blown around in
dust and spreads quite readily. Some lizards sicken and die from it,
others can carry it. If one lizard in the house has it, very likely, all
may have the virus. This virus resides in tissue, and will be shed in
varying rates. All of this makes the virus a hard one to pin down,
prevent or study.

About our CMI Staff
The CMI lab is run on a day-to-day basis by Lou Ann Miller MT(ASCP)
under the Administration and supervision of Dr. Edwin Hahn, Associate
Dean for Research and Advanced Studies.

Lou Ann is a board certified Medical Technologist by the board of the
American Society of Clinical Pathology, and a board certified Electron
Microscopist by the Microscopy Society of America.



Thank you Lou Ann for your wonderful article of information.  Your contribution is invaluable to us.