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question for BSE tester

cedardell

Well-known member
Would be feasable to use your urine test in the private sector? Could a veterinarian or a rancher use it? If so what would it cost to get set up to use it and how would you go about it?
 

bse-tester

Well-known member
First of all the problem with setting up in the private sector is funding and that is hotly followed by licensing. The license to test is traditionally given by the very authorities that want to conduct the testing so it becomes a vicious circle of red tape. The only way to do it is to bring a large and overwhelming amount of ranchers to the table and lobby the authorities for a license to first of all, get inot conducting a "Pilot Project." Heck, I would even suggest getting government funding to do that. Once you prove that it will work in the proivate sector, approach the government and hammer them with your data.

A rancher could NOT use it - due to the fact that we then have what is known as the fox looking after the chickens. Most ranchers simply do not have the time, let alone the training to conduct the test and on top of this, they will certainly not have a laboratory that is set up to conduct prion disease research in. The cost of the lab will be in the region of US$250 - 500,000.00 and that is before you hire the technical staff to run it.
 

PORKER

Well-known member
The license to test is traditionally given by the very authorities that want to conduct the testing so it becomes a vicious circle of red tape.

The womans preg test gets cheaper and easier to use every day. Too bad the BSE live animal test couldn't be this easy.
 

Jason

Well-known member
A woman's pregnancy test and a BSE test are 2 very differnt matters.

I doubt anywhere on the planet you will find a government that allows food companies to set testing procedures for disease and safety.

If a woman gets a false positive with a pregnancy test, she usually goes to her doctor and gets it checked. Other than a slight scare or a let down depending if she wants to be pregnant or not, no harm done.
 

Mike

Well-known member
Jason said:
A woman's pregnancy test and a BSE test are 2 very differnt matters.

I doubt anywhere on the planet you will find a government that allows food companies to set testing procedures for disease and safety.

If a woman gets a false positive with a pregnancy test, she usually goes to her doctor and gets it checked. Other than a slight scare or a let down depending if she wants to be pregnant or not, no harm done.

Are you conceding that a BSE test is a FOOD SAFETY test?

HAACCP allows companies to regulate themselves on food safety tests everyday. They also set the recall procedures.
 

Econ101

Well-known member
Jason said:
A woman's pregnancy test and a BSE test are 2 very differnt matters.

I doubt anywhere on the planet you will find a government that allows food companies to set testing procedures for disease and safety.

If a woman gets a false positive with a pregnancy test, she usually goes to her doctor and gets it checked. Other than a slight scare or a let down depending if she wants to be pregnant or not, no harm done.

And a positive bse test should have similar effects, with oversight from an honest government.

Jason, when if ever are you going to start promoting the truth?
 

Jason

Well-known member
I don't promote bogus testing. Testing young animals, or using unproven tests.

I don't think BSE is a food safety issue based on what is known about it. About 150 people have been claimed to contract it from eating beef, but beef without SRM removal.

Based on that BSE is an animal health issue, but SRM removal is a food safety issue. And yes sometimes one affects the other.

The ones not promoting truth are those that want unproven tests used on animals too young to show BSE. USDA could make 35 million cattle under 30 months pee on a stick, call it 100% testing and still never find a case.

That is promoting a lie.
 

Econ101

Well-known member
Jason said:
I don't promote bogus testing. Testing young animals, or using unproven tests.

I don't think BSE is a food safety issue based on what is known about it. About 150 people have been claimed to contract it from eating beef, but beef without SRM removal.

Based on that BSE is an animal health issue, but SRM removal is a food safety issue. And yes sometimes one affects the other.

The ones not promoting truth are those that want unproven tests used on animals too young to show BSE. USDA could make 35 million cattle under 30 months pee on a stick, call it 100% testing and still never find a case.

That is promoting a lie.

Who is promoting bogus testing? I want the tests to be validated, if they can be. If they can not be validated, then that should be known through an efficient validation process.

If we don't have a process that works for validating tests, how can we know if our tests (any tests) are accurate? They are due for manipulation.

Who made you a bse testing expert, Jason? It seems to me not too long ago you were an expert on corn production.
 

Mike

Well-known member
The ones not promoting truth are those that want unproven tests used on animals too young to show BSE. USDA could make 35 million cattle under 30 months pee on a stick, call it 100% testing and still never find a case.

This is your opinion and your opinion only. The issue is not whether the urine test is valid or not. (It does include Western Blot Technology-which has certainly been validated)

But the issue is whether the USDA wants to validate it or not. From all we can assess from the situation is that they do not want to validate nor invalidate it.

If technology quits moving forward we will be stuck with the BSE/cattle scare for eternity. Do you not want to improve demand?
 

Jason

Well-known member
Did you not read the article OT posted about the urine test?

It was very clear the test is still experimental. It is not ready for validation. It still needs to be tested to assure it even works.

Promoting a test that is still experimental with a disease that is still not fully understood isn't in anyone's best interest.

If the test shows promise, there might come a time when the USDA and or CFIA could include a validation process where the urine test is conducted beside a proven test. That time isn't here.

Even if the urine test does turn out to work, it is not a garantee that a young animal that is still in the incubation stage of BSE will trigger a positive.

The risk is testing an animal at too young of an age, then it develops misfolded prions later in life, but carries a BSE free tag.
 

Econ101

Well-known member
Jason said:
Did you not read the article OT posted about the urine test?

It was very clear the test is still experimental. It is not ready for validation. It still needs to be tested to assure it even works.

Promoting a test that is still experimental with a disease that is still not fully understood isn't in anyone's best interest.

If the test shows promise, there might come a time when the USDA and or CFIA could include a validation process where the urine test is conducted beside a proven test. That time isn't here.

Even if the urine test does turn out to work, it is not a garantee that a young animal that is still in the incubation stage of BSE will trigger a positive.

The risk is testing an animal at too young of an age, then it develops misfolded prions later in life, but carries a BSE free tag.

Jason, just admit you don't know what you are talking about.
 

bse-tester

Well-known member
Jason wrote:

Did you not read the article OT posted about the urine test?

It was very clear the test is still experimental. It is not ready for validation. It still needs to be tested to assure it even works.

Promoting a test that is still experimental with a disease that is still not fully understood isn't in anyone's best interest.

If the test shows promise, there might come a time when the USDA and or CFIA could include a validation process where the urine test is conducted beside a proven test. That time isn't here.

Even if the urine test does turn out to work, it is not a garantee that a young animal that is still in the incubation stage of BSE will trigger a positive.

The risk is testing an animal at too young of an age, then it develops misfolded prions later in life, but carries a BSE free tag.

Jason, with all due respect, the statment you made, as quoted about, is about as ridiculous a comment as I have ever read on this board.

The test does work! It is ready for validation! and it will be validated - period!! The test, like any other test that is "Pre-validation" is considered experimental. That is simply a use of words. The test was proven in the US National Laboratories and is now ready to validate.

As for testing young animals - age is NOT a factor and the test simply doesn't care how old the animal was at the time of taking the urine sample!! If the disease is in the animal, it will be detected as all cattle, like humans, of any age, excrete prions. Get your facts straight before you write your comments.

Now you can say that you heard it from one who truly does know what the hell he is talking about!!!
 

bse-tester

Well-known member
Murgen wrote:

Has anyone ever tested feed and found mis-folded prions?

The British Government conducted a series of tests on feed during the BSE crisis over there and found lots of contamination from PrPsc.
 

flounder

Well-known member
USDA Testing Protocols and Quality Assurance Procedures

In November 2004, USDA announced that its rapid screening test produced an inconclusive BSE test result. A contract laboratory ran its rapid screening test on a brain sample collected for testing and produced three high positive reactive results. As required, the contract laboratory forwarded the inconclusive sample to APHIS’ National Veterinary Services Laboratories (NVSL) for confirmation. NVSL repeated the rapid screening test, which again produced three high positive reactive results. Following established protocol, NVSL ran its confirmatory test, an immunohistochemistry (IHC) test, which was interpreted as negative for BSE.

Faced with conflicting results between the rapid screening and IHC tests, NVSL scientists recommended additional testing to resolve the discrepancy but APHIS headquarters officials concluded that no further testing was necessary since testing protocols were followed and the confirmatory test was negative. In our discussions with APHIS officials, they justified their decision to not do additional testing because the IHC test is internationally recognized as the "gold standard" of testing. Also, they believed that

USDA/OIG-A/50601-10-KC/ Page iv

conducting additional tests would undermine confidence in USDA’s testing protocols.

OIG obtained evidence that indicated additional testing was prudent. We came to this conclusion because the rapid screening tests produced six high positive reactive results, the IHC tests conflicted, and various standard operating procedures were not followed. Also, our review of the relevant scientific literature, other countries’ protocols, and discussions with experts led us to conclude that additional confirmatory testing should be considered in the event of conflicting test results.

To maintain objectivity and independence, we requested that USDA’s Agricultural Research Service (ARS) perform the Office International des Epizooties (OIE) Scrapie-Associated Fibrils (SAF) immunoblot test. The additional testing produced positive results. To confirm, the Secretary of Agriculture requested that an internationally recognized BSE laboratory in Weybridge, England (Weybridge) perform additional testing. Weybridge conducted various tests, including their own IHC tests and three Western blot tests. The tests confirmed that the cow was infected with BSE. The Secretary immediately directed USDA scientists to work with international experts to develop new protocols that include performing dual confirmatory tests in the event of an inconclusive BSE screening test.

We attribute the failure to identify the BSE positive sample to rigid protocols, as well as the lack of adequate quality assurance controls over its testing program. Details of our concerns are discussed in Findings 3 and 4.



snip...



Section 2. Testing Protocols and Quality Assurance Controls In November 2004, USDA announced that its rapid screening test, Bio-Rad Enzyme Linked Immunosorbent Assay (ELISA), produced an inconclusive BSE test result as part of its enhanced BSE surveillance program. The ELISA rapid screening test performed at a BSE contract laboratory produced three high positive reactive results.40 As required,41 the contract laboratory forwarded the inconclusive sample to the APHIS National Veterinary Services Laboratories (NVSL) for confirmatory testing. NVSL repeated the ELISA testing and again produced three high positive reactive results.42 In accordance with its established protocol, NVSL ran its confirmatory test, an immunohistochemistry (IHC) test, which was interpreted as negative for BSE. In addition, NVSL performed a histological43 examination of the tissue and did not detect lesions44 consistent with BSE. Faced with conflicting results, NVSL scientists recommended additional testing to resolve the discrepancy but APHIS headquarters officials concluded no further testing was necessary because testing protocols were followed. In our discussions with APHIS officials, they justified their decision not to do additional testing because the IHC is internationally recognized as the “gold standard.” Also, they believed that conducting additional tests would undermine confidence in USDA’s established testing protocols. However, OIG obtained evidence that indicated additional testing was prudent to ensure that USDA’s testing protocols were effective in detecting BSE and that confidence in USDA’s testing procedures was maintained. OIG came to this conclusion because the rapid tests produced six high positive reactive results, confirmatory testing conflicted with the rapid test results, and various standard operating procedures were not followed. Also, our review of scientific literature, other country protocols, as well as discussions with internationally recognized experts led us to conclude that confirmatory testing should not be limited when conflicting test results are obtained. To maintain objectivity and independence in our assessment, we requested the USDA Agricultural Research Service (ARS) perform the Office International des Epizooties (OIE) Scrapie-Associated Fibrils (SAF) 40 ELISA test procedures require two additional (duplicate) tests if the initial test is reactive, before final interpretation. If either of the duplicate tests is reactive, the test is deemed inconclusive. 41 Protocol for BSE Contract Laboratories to Receive and Test Bovine Brain Samples and Report Results for BSE Surveillance Standard Operating Procedure (SOP), dated October 26, 2004. 42 The NVSL conducted an ELISA test on the original material tested at the contract laboratory and on two new cuts from the sample tissue. 43 A visual examination of brain tissue by a microscope. 44 A localized pathological change in a bodily organ or tissue.

immunoblot.45 ARS performed the test at the National Animal Disease Center because NVSL did not have the necessary equipment46 (ultracentrifuge) to do the test. APHIS scientists observed and participated, as appropriate, in this effort. The additional tests conducted by ARS produced positive results. To confirm this finding, the Secretary requested the internationally recognized BSE reference laboratory in Weybridge, England, (Weybridge) to perform additional confirmatory testing. Weybridge conducted various tests, including their own IHC methods, as well as three Western blot methods. The tests confirmed that the suspect cow was infected with BSE. Also, Weybridge confirmed this case as an unequivocal positive case of BSE on the basis of IHC. As a result of this finding, the Secretary immediately directed USDA scientists to work with international experts to develop a new protocol that includes performing dual confirmatory tests in the event of another inconclusive BSE screening test. Finding 3 Rigid Protocols Reduced the Likelihood BSE Could be Detected APHIS relied on a single test method, as well as a histological examination of tissue for lesions consistent with BSE, to confirm the presence of BSE even though discrepant test results indicated further testing may be prudent. When IHC test results were interpreted as negative, APHIS concluded the sample tested negative for BSE. Subsequent independent tests initiated by OIG using a different testing method, as well as confirmatory testing by Weybridge, determined that the suspect sample was a positive case of BSE. APHIS Declares BSE Sample Negative Despite Conflicting Results When the tissue sample originally arrived at NVSL in November 2004 from the contract lab, NVSL scientists repeated the ELISA screening test and again produced three high positive reactive results. NVSL scientists cut out two sections of the brain sample for IHC testing. One section was used for an experimental procedure that was not part of the confirmatory testing protocol, and the other cut was for normal IHC testing using scrapie for a positive control.47 According to NVSL scientists, the experimental test results were inconclusive but the IHC test was interpreted as negative. The NVSL scientists were concerned with the inconsistencies and conducted 45 The OIE SAF immunoblot is an internationally recognized confirmatory test, often referred to as a Western blot test. There are different types of Western blots; the OIE SAF immunoblot includes enrichment steps taken with the sample prior to the standard Western blot steps. 46 APHIS has now ordered the necessary equipment for NVSL. USDA/OIG-A/50601-10-KC Page 32

47 A positive control is a sample that is known to contain a given disease or react in the test. The sample then can be used to make sure that the test for that disease works properly. In the case of BSE, tissue infected with either scrapie or BSE can serve as a positive control for an IHC test for BSE since both are different forms of the same disease (transmissible spongiform encephalopathy or TSE).

another IHC test using BSE as a positive control.48 The test result was also interpreted as negative. Also, according to the NVSL scientists, the histological examination of the tissue did not detect lesions consistent with BSE. After the second negative IHC test, NVSL scientists supported doing additional testing. They prepared a plan for additional tests; if those tests had been conducted, BSE may have been detected in the sample. The additional tests recommended by NVSL scientists, but not approved by APHIS Headquarters officials, were the IHC using other antibodies (IHC testing using different antibodies ultimately produced positive results); IHC testing of additional regions of the brain (the cerebellum tested positive); regular and enriched (OIE-like) Western blots (the obex and cerebellum tested positive); and variable rapid tests (the obex and cerebellum tested positive with two different rapid tests). NVSL officials also recommended that the sample be sent to Weybridge for confirmatory testing (to conduct IHC and OIE Western blot tests). In June 2005, Weybridge conducted IHC testing with three different antibodies, including the antibody used in the United States (tested positive), the OIE Western blot (tested positive), a modified commercial kit Western blot (negative) and the NaPTA49 Western blot (tested positive). We obtained information as to the differing protocols used by other countries. We found that while APHIS determined that additional testing was unnecessary after the IHC test, other countries have used multiple tests to confirm positives. In Japan, for example, all reactive screening test samples are examined by both IHC and a Western blot (different from the OIE SAF immunoblot). In the United Kingdom (U.K.), IHC and Western blot (different from the OIE SAF immunoblot) tests are used for all animals that test positive with a screening test. When IHC and the Western blot fail to confirm a positive rapid test, the U.K. resorts to a third test, the OIE SAF immunoblot. With these procedures in place, both Japan and the U.K. have found BSE cases that were rapid test reactive, IHC negative, and finally confirmed positive with a Western blot. Evidence Indicated Additional Testing Would Be Prudent We also spoke with an internationally recognized BSE expert regarding the advisability of limiting confirmatory testing when conflicting results are obtained. This official expressed concern about limiting confirmatory tests to the IHC despite its status as one of the “gold standard” tests. He advised that the IHC is not one test; it is a test method that can vary significantly in sensitivity from laboratory to laboratory. New antibodies can improve or

USDA/OIG-A/50601-10-KC Page 33

48 The NVSL uses scrapie as the positive control as part of its normal IHC testing procedures. Due to the conflicting results between the ELISA and IHC tests, the NVSL conducted another IHC test with BSE as the positive control. Subsequently, the NVSL modified the Confirming Inconclusive Results from BSE Testing Laboratories at the NVSL SOP to show that all IHC tested BSE inconclusive samples from contract laboratories will use BSE as the positive control. 49 Sodium phosphotungstic acid.

USDA/OIG-A/50601-10-KC Page 34

reduce sensitivity, as can variations in many of the reagents50 used. He explained that his laboratory had experienced cases where an initial confirmatory IHC test was challenged by either a more extensive IHC test or “…applying a more sensitive immunoblot.” He emphasized the importance of having additional confirmatory testing to resolve discrepant results since there are many variables, and most of the variability appears to be due to test performance of the laboratory. OIG became concerned that APHIS relied on its confirmatory test methods when rapid screening tests produced high positive reactive results six times.51 Also, we found that APHIS did not pursue and/or investigate why the ELISA produced high reactive positives. An official from the manufacturer of the ELISA test kit told us that they requested, but did not receive, information on the inconclusive reported by USDA in November 2004. These officials requested this information in order to understand the reasons for the discrepant results. The Bio-Rad ELISA rapid screening test is internationally recognized as a highly reliable test and is the rapid screening test used for USDA’s surveillance effort. According to APHIS officials, they felt it would be inappropriate to collaborate on the one sample because Bio-Rad is a USDA-APHIS regulated biologics company and only one of several competing manufacturers. To maintain confidence in USDA’s test protocols, it would have been a prudent course of action for USDA to determine why such significant differing results were obtained. The fact that they did not pursue this matter caused concerns relating to testing quality assurance procedures. In this regard, we found lack of compliance with SOPs relating to laboratory proficiency and quality assurance (see Finding 4), and, in this case, the storage of sampled material and reporting of test results. We found that the NVSL did not prepare a report to document its confirmatory testing of the November 2004 sample. The SOP52 states that the BSE network laboratory initiating the inconclusive will receive a report of the case. NVSL officials could not explain why a final report had not been prepared. We also found that the inconclusive sample was frozen prior to IHC confirmatory testing. APHIS protocols state that samples are not to be frozen prior to laboratory submission. The OIE Manual of Diagnostic Tests and Vaccines for Terrestrial Animals states that the tissues for histological or IHC examination are not to be frozen as this will provide artefactual53 lesions that may compromise the identification of vacuolation,54 and/or target site location. Although the sample was frozen, APHIS did not conduct a Western 50 A substance used in a chemical reaction to detect, measure, examine, or produce other substances. 51 The six high positive reactive results were from three tests of the submitted sample (multiple runs of the same test). 52 Confirming Inconclusive Results from Bovine Spongiform Encephalopathy Testing Laboratories at the NVSL SOP, dated August 13, 2004. 53 A structure or feature not normally present but visible as a result of an external agent or action, such as one seen in a microscopic specimen after fixation. 54 A small space or cavity in a tissue.

USDA/OIG-A/50601-10-KC Page 35

blot test on the sample. An NVSL official said freezing the sample does not make it unsuitable for IHC. APHIS determined that the sample was suitable for IHC and therefore, in accordance with its SOP, did not conduct a Western blot test. APHIS also handled the December 2003 BSE positive differently than the November 2004 sample. For the December 2003 BSE positive sample, APHIS conducted several confirmatory tests in addition to the IHC testing and histological examination (unlike the November 2004 sample tests, both of these were interpreted as positive). ARS performed two Western blots (Prionics Check Western blot and an ARS developed Western blot). When we questioned why the samples were handled differently, APHIS officials stated that the Western blots were done because the IHC in December 2003 was positive. The additional testing was done to further characterize the case, because it was the first U.S. case; the additional testing was not done to decide whether the case was positive or negative. We discussed our concerns with limiting confirmatory testing, particularly given conflicting results, with the APHIS Administrator and staff in May 2005. He explained that international standards recognized more than one “gold standard” test. In setting up its testing protocols, USDA had chosen one as the confirming test, the IHC test, and stayed with it. APHIS protocols only allow a Western blot in cases where the sample has become unsuitable for IHC tests (e.g., in cases where the brainstem architecture is not evident). International standards, he continued, accept a tissue sample as negative for BSE if its IHC test is negative. Once the test is run in accordance with protocols, additional tests undermine the USDA testing protocol and the surveillance program. He concluded that since APHIS’ protocols accepted the IHC test as confirming the presence or absence of BSE, no further testing was necessary. According to protocol, the tissue sample was determined to have tested negative for BSE. On June 24, 2005, USDA announced that the additional testing by the BSE reference laboratory in England confirmed the presence of BSE in the tissue sample. To obviate the possibility that a future sample would be declared negative and then found positive, the Secretary of Agriculture announced a change to APHIS’ testing protocols that same day. He called for “dual confirmatory tests in the event of another ‘inconclusive’ [reactive] BSE screening test.” He also indicated that he would reinforce proper procedures so that samples will not be frozen, and to spot-check the laboratories to see that they complete reports as required. APHIS issued a SOP on the new confirmatory testing protocols on November 30, 2005.



http://www.usda.gov/oig/webdocs/50601-10-KC.pdf



TSS
 

Econ101

Well-known member
Also, they believed that conducting additional tests would undermine confidence in USDA’s established testing protocols.


and they were right!!!!
 
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