The Alchemy of Flow Cytometry
FROM RESEARCH TO JUNK SCIENCE
In 1972, Congress established the Office of Technology Assessment (
OTA) to serve the legislative branch as an independent source of information and analysis about complex scientific and technical issues. OTA construed health technology broadly, including “all elements of medical practice that are knowledge-based, including hardware (equipment and facilities) and software (knowledge skills)… the set of techniques, drugs, equipment, and procedures used by health care professionals in delivering medical care to individuals and the systems within which such care is delivered.”
By 1978, the OTA produced a shattering report on the state of scientific medicine,
Assessing the Efficacy and Safety of Medical Technologies. The report stated:
“Evidence indicates that many technologies are not adequately assessed before they enjoy widespread use… Many technologies which have been used extensively have later been shown to be of limited usefulness”…and ” … only 10 to 20 percent of all procedures currently used in medical practice have been shown to be efficacious by controlled trial.”
The report implied that 80% to 90% of all routinely-performed procedures are unproven – a conclusion that implicates the technology of flow cytometry that uses immunophenotyping to identify antigen markers on various cell populations.
The
U.S. News and World Report issue of 23 November 1987 raised further questions about HIV tests:
“With public health officials and politicians thrashing out who should be tested for HIV, the accuracy of the test itself has been ignored. A study last month by the Congressional Office of Technology Assessment found that HIV tests can be very inaccurate indeed. For groups at very low risk – people who do not use IV drugs or have sex with gay or bisexual men – 9 in 10 positive findings are called false positives, indicating infection where none exists.”
OTA’s warning continues to be ignored by the medical and scientific communities and the politicians, agencies and regulators that enable them.
In 1996, Congress disbanded the OTA, leading to the systematic deregulation of the various medical technology industries. The U.S. Supreme Court sealed the fate of future scientific transparency by ruling in favor or corporate interests in the
Citizens United v. Federal Election Commission (2010).
The OTA’s demise closed a low-budget item that gave Americans too much information to make informed choices. It paved the way for the establishment of a medical and scientific knowledge monopoly that is now permeated by corruption and fraud (junk science).
As a result, the “AIDS industry” continues to use unproven technologies like flow cytometry to diagnose, alarm, and treat healthy people with toxic and expensive drugs that are
designed to make them sick (see
video).
Fundamental issues regarding the limitations of flow cytometry technology and the propriety of its use in the diagnosis of acquired immune deficiency have never been addressed:
- Proofthat an exogenous virus uniquely attacks CD4+T cells and is cytopathic (see above discussion). Further, the relationship of the Th1/Th2 balance shift in the CD4+T cell population has been ignored.
- Reproducibility – samples drawn concurrently from one subject should deliver the same results in every machine and laboratory that receives those specimens.
Police officers have successfully used
gas chromatography (GC) in the enforcement of drunk driving laws for many years. When properly operated, a functional and calibrated
BreathalyserTM not only measures blood alcohol levels in breath samples accurately (sensitivity), but they can reliably differentiate between subjects that have –
and have not – consumed alcohol (specificity).
Police officers also use RADAR to enforce basic speed laws. But like GC, officers do not rely upon RADAR devices to enforce laws. Instead, officers rely upon their training and expertise to estimate intoxication and velocities. Once they develop articulable facts that indicate impairment or an unsafe speed, they use GC and RADAR
to confirm what their training, expertise and observations initially tell them.
Unlike GC and RADAR, flow cytometry manufacturers compensate for their inaccuracies by inventing proprietary algorithms to report spurious and unreliable cell counts. There is no reliable method for counting standardization for products and operators, and the substantial deviations in technical competency and quality control of test samples between labs render these tests wholly unreliable. Clinicians simply order blood draws and
presume that lab results are accurate and meaningful. Clinicians who receive kickbacks from labs and drug companies have little incentive to question results of their asymptomatic patients.
This protocol is akin to policemen who stop 35 mph motorists because their RADAR device captured a 90mph reading. But while police agencies would train or terminate such derelict employees, this practice – when applied to biological testing and flow cytometry – is considered the “medical standard of care.”
Flow cytometry employs two techniques to count cells:
- Dual Platform Systems – One component determines cell concentrations, while the second determines the relative number of CD4 and CD8 cells. Unless these two components count a common parameter, dual platform systems cannot accurately correlate the results.
- Single Platform Systems – These platforms are especially designed to count the absolute numbers of antibody-labeled cells. These devices are equipped with multiple sample loader, programming facility and computer support, which removes the need for using two different machine to determine the concentration of CD4 and CD8 cells.
The variability of results when comparisons are made between these two counting systems has been shown to be as
high as 56%.
- Standardization – HHS, CDC, NIH and FDA have failed to produce meaningful guidelines for quality control, quality assurance or quality of test reagents.
Flow cytometric immunophenotyping is a relatively new technology that is highly complex, involves multiple components and procedures, and has numerous points for measurement vulnerability. As the technology moved from research laboratories to clinical laboratories, the need for standardization increased. In response to that need, guidelines addressing aspects of the CD4+T lymphocyte testing process – in particular, quality control, quality assurance, and consistency of reagents for immunophenotyping of lymphocytes were developed. (National Institute of Allergy and Infectious Diseases (NIAID)/AIDS Clinical Trial Group: Guidelines for hematologic and low cytometric analysis of ACTG specimens, 1992).
To assure the accuracy and reliability of CD4+T cell test results obtained within individual laboratories and to attempt to assure comparability of results between laboratories, the CDC established a list of standard methods for performing the test, as well as guidelines for quality control and quality assurance. The CDC’s recommendations for flow cytometry apply to laboratory safety, specimen collection, specimen transport, maintenance of specimen integrity, specimen processing, flow cytometer quality control, sample analyses, data analysis, data storage, data reporting and
quality assurance.
As can be seen, there are multiple steps in this process, any of which that if violated can lead to a substantial alteration in the test results:
1.
Blood collection: The type of vial, the time of day and the temperature at which the specimen is handled can all have an effect on test results. CD4+T cell counts are known to be higher in the afternoon than in the morning – a result of the response to the diurnal variation in steroid production from the adrenal gland.
2.
Specimen transport: Was the specimen maintained at room temperature? If the specimen is too hot or too cold, cellular destruction might occur. This can be a major problem for transporting of the specimen outside of the collection facility.
3.
Specimen Integrity: When the specimen was received, was it too hot or too cold? Was the blood hemolyzed or frozen? Are there visible clots? Has the specimen been received > 72 hours after collection? If so, the specimen must be rejected.
4.
Specimen processing: The test should be run within 48 hours, but no later than 72 hours after drawing the blood. Procedures that must be followed:
5. Gently rocking blood for 5 minutes to ensure uniform sample
6. Pipetting accurate blood volumes; vortex sample tubes to mix the blood and reagents and break up cell aggregates
7. Quality and type of reagent used
8. Incubating tubes in dark during staining procedure
9. A lyse/no wash method which requires following manufacture directions (each manufactures has a different set of directions and a different counting algorithm)
10. An immediate capping and storing all stained samples in the dark under refrigeration until flow cytometric analysis is performed.
11. It is advised that the specimens be stored no more than 24 hours unless the laboratory can demonstrate that scatter and fluorescence patterns do not change for specimens stored for longer periods.
12.
Machine calibration: Variations in absolute lymphocyte counts obtained by different automated cell counters exposes another problem. A review of
four widely used automated counters indicate that analytic variability in the absolute lymphocyte counts due primarily to method variability, is significant and larger than the variability typically observed on inter-laboratory trials of relative CD4+T cell counts. These method biases cannot easily be reduced by calibration, since
the cell classification algorithms are built in features of the various counters.
As can be seen, the process of flow cytometry using immunophenotyping requires not only a sophisticated level of technical skill, but a chain of delivery and processing events that is probably difficult to replicate from lab to lab, but also to substantiate. One study found
errors of 18% and 35% of absolute CD4+T cell count, while another study found the inter-laboratory variability so significant that it led to
conflicting treatment recommendations.
continued...
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