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Subject: [WNV-L] Efficacy of VecTest for WNV Detection

Date: July 3, 2003
Posted by: Lois Levitan (lcl3@cornell.edu)
Ward Stone (wbstone@gw.dec.state.ny.us)
Kirti Dave (kdave@mas-inc.com)


The following is a series of correspondence between Ward Stone, 
Wildlife Pathologist with the NYS Department of Environmental 
Conservation, and Kirti DavŽ, Team Director at Medical Analysis 
Systems, Inc., which developed the VecTest.   The first memo [1] was 
written to me, in response to my request for permission to post the 
correspondence to the WestNileVirus-L listserv (since I had been 
included on the exchange as an individual not as listserv moderator). 
Other memos are in the order they were written, from earliest to most 
recent.  -- Lois Levitan


[1]  Permission to post
[2]  Problems with West Nile Virus VecTests.  June 10, 2003.  Ward 
Stone to Kirti Dave
[3]  Response to "Problems." June 10, 2003. Kirti Dave to Ward Stone
[4] Cautions on use of the VecTest for West Nile Virus diagnosis in 
birds. July 1, 2003. Ward Stone to Kirti Dave and recipient list
[5]  Response to "Cautions on use of VecTest." July 1, 2003. Kirti 
Dave to Ward Stone and recipient list.

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[1]  PERMISSION TO POST
Date: Thu, 03 Jul 2003 12:51:49 -0400
From: Ward Stone by way of "Joseph Therrien" <jetherri@gw.dec.state.ny.us>
To: <lcl3@cornell.edu>

You can post our data on the VecTest on the West Nile Virus-L 
listserv. I am also transmitting our previous email from my unit to 
and from Dr. Dave. I had hoped that organ tissues (e.g. brain, 
kidney, liver, spleen) would show more accuracy and precision for the 
presence of WNV antigen than the oral epithelium. This does not seem 
to be the case, but the oral swabs also show a significant false 
positive and false negative problem - even in corvids.

Dr. Ward B. Stone M.S., sc. D.
Wildlife Pathologist
NYS Department of Environmental Conservation
Wildlife Pathology Unit
108 Game Farm Rd
Delmar, NY 12054

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[2]  PROBLEMS WITH THE WEST NILE VIRUS VECTESTS

>From: Ward Stone [mailto:wbstone@gw.dec.state.ny.us]
>Sent: Tuesday, June 10, 2003 12:56 PM
>To: kdave@mas-inc.com
>Subject: Problems with West Nile Virus VecTests
>
>In testing the VecTest  West Nile Virus Antigen Assay that your 
>company supplied to the Wildlife Pathology Unit of the New York 
>State DEC, we have found a number of problems with false positives. 
>Although we will be carrying out extensive further testing, the data 
>we have to date causes me not to use your companies VecTests for 
>diagnosing the presence of West Nile in birds - including crows.
>
>Are other laboratories/scientists experiencing similar difficulties 
>with the VecTest?
>
>Attached is a summary of our data as to noon today. I am especially 
>concerned that people may  be using at least some test kits that 
>your company has supplied that are unreliable.
>
>Ward B. Stone   BA, MS, SC.D
>Wildlife Pathologist II
>New York State Department of Environmental Conservation
>Wildlife Pathology Unit
>108 Game Farm Road
>Delmar, New York 12054
>Phone (518) 478-3032
Fax (518) 478-3035

-----

[3]  RESPONSE TO "PROBLEMS WITH THE WEST NILE VIRUS VEC TEST"

>From: Kirti Dave <kdave@mas-inc.com>
>To: 'Ward Stone' <wbstone@gw.dec.state.ny.us>
>Subject: RE: Problems with West Nile Virus VecTests
>Date: Tue, 10 Jun 2003 14:32:25 -0700
>
>Dr Stone,
>
>The VecTest kits/assays are not intended to be used on any tissue 
>(tissue extracts). From the results you sent, amongst all samples 
>tested there are two oral swabs out of which only one of them is 
>from a corvid that has
>tested positive. For this sample I would ask you to repeat the PCR 
>and VecTest again. Also it would be worth while to check out with 
>another antigen based assay such as ELISA.
>
>The last sample from Westchester which tested positive by PCR but 
>negative with VecTest, was that a marginal virus amount?
>
>The test kits we market presently are for the use with mosquito 
>samples only. Do you test these? The use of VecTest WNV Antigen 
>assay for oral swabs collected from corvids appears to be working 
>for labs that have initiated these type of use. The problems that 
>would come up would be when the kits become a tool for any kind of 
>exploratory work. The reagents used in the dipstick assay are 
>monoclonals and to avoid nonspecific interactions with IgGs from 
>other species and/or other components, one would need to modify the 
>current format of the dipstick.
>
>The data presently at the AMCA March, 2003, from oral swabs from 
>corvids is good enough to continue using the VecTest for that 
>purpose. This data is still being collected so that we can market a 
>kit for that purpose. Regarding cautions from findings about 
>exploratory uses such as tissues and other species, we will be open 
>and point that out to the customers. The kits were never meant to 
>work in every and any sample.
>
>I would be grateful if you provide me more quantitative data on 
>corvid oral samples where VecTest  provided a result different from 
>the PCR. That data would have meaning for other customers and for 
>something to check out for scientific interest and a just evaluation 
>of a new rapid assay tool. I hope
>you will be open and continue to provide me feedback.
>
>Thanking you,
>Best Regards,
>
>Kirti
>
>Kirti DavŽ PhD
>Senior Project Manager
>Medical Analysis Systems, Inc
>5300 Adolfo Road
>Camarillo, CA 93012
>Phone: (805)-987-7891 ext 6-3214
Fax: (805)-383-8260

-----

[4]  CAUTIONS ON USE OF THE VECTEST FOR WEST NILE VIRUS DIAGNOSIS IN BIRDS

>From: Ward Stone by way of Joseph Therrien 
>[mailto:jetherri@gw.dec.state.ny.us]
>Sent: Tuesday, July 01, 2003 1:47 PM
>To: nap4@cdc.gov; nck6@cdc.gov; lcl3@cornell.edu; Roscoe@eclipse.net;
>campbell@gw.dec.state.ny.us; ldk02@gw.dec.state.ny.us;
>mxe04@gw.dec.state.ny.us; ebk01@health.state.ny.us; kdave@mas-inc.com;
>doug docherty@usgs.gov; esaito@usgs.gov; kathy_converse@usgs.gov
>Subject: VecTest Results
>
>Cautions on use of the VecTest for West Nile Virus diagnosis in birds.
>
>The attached table* shows the frequency of false positives for WNV 
>in birds tested to date with the VecTest. Although oral false 
>positives are relatively low in percentage, they nevertheless are 
>significant. From the literature we had not expected this level of 
>false positives in the oral tests. False positives for WNV were 
>common in the tests run on kidney and liver samples. At this point 
>New York has only two birds that
>are WNV positive by PCR. Both of these are crows with false 
>negatives by VecTest.
>
>As the season progresses we anticipate numerous WNV infected birds 
>to arrive at our lab and this will provide many cases, of a variety 
>of species, to check for false negatives.
>
>Our data indicates that the use of the VecTest should be used with 
>caution and that the VecTest seems to need more adjustment and/or 
>development before it is fully acceptable as a diagnostic aid for 
>WNV in birds.
>
>Dr. Ward B. Stone
>Wildlife Pathologist
>Wildlife Pathology Unit
>108 Game Farm Road
>Delmar, Ny 12054
>
>Phone (518) 478-3032
Fax (518) 478-3035

* See 
http://environmentalrisk.cornell.edu/WNV/WNVEducDocs/VecTestTissueResults.xls

---

[5]  RESPONSE TO "VECTEST RESULTS"

Date: Tue, 1 Jul 2003 16:13:40 -0700
From: Kirti Dave <kdave@mas-inc.com>
To: [recipient list for Ward Stone's message]
Subject: RE: VecTest Results


Dr Ward Stone,

Thank you for the data. Two things to point out:

(1) Use of VecTest antigen detection assay in detection of virus 
antigen in tissue samples has been carried out on purely exploratory 
basis in this study. This is not recommended and emphatically stated 
so earlier (my
correspondence dated 6/10/03 attached herewith).

(2) It would be worthwhile to figure out why an antigen detection 
test such as VecTest would be positive or negative in the oral swabs 
where its best use could be - in rapid dead bird analysis or 
surveillance area. Any
quantitative data on the concentration of the virus in the 4+1 corvid 
oral samples that were falsely positive and the two samples you 
mentioned that are false negatives?

The part of the data where the test is used properly such as on oral 
swabs in your study (attached herewith) is good information and when 
supported with quantitative data could be used for determining 
caution in use for oral swab analysis of corvid/non corvid species.

And again I want to reassure you and others copied on this email that 
if the data from  proper use does not support usefulness of the 
product, we will discontinue providing the product for that use. We 
want to provide the best and reliable tools. Therefore feel free to 
provide feedback.

Regards,

Kirti

Kirti DavŽ PhD
Team Director
Medical Analysis Systems, Inc
5300 Adolfo Road
Camarillo, CA 93012
Phone: (805)-987-7891 ext 6-3214
Fax: (805)-383-8260
email: kdave@mas-inc.com


-- 
Lois Levitan, PhD       Program Leader
Environmental Risk Analysis Program
Department of Communication and
Center for the Environment
213 Rice Hall, Cornell University
Ithaca, New York USA 14853-5601

Phone:   (607) 255-4765     Fax: (607) 255-0238
Email:	LCL3@cornell.edu
Web:       http://environmentalrisk.cornell.edu
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