Session 7102 You Want to Measure “That” in “This”? Body Fluid Testing in the Clinical Laboratory Academy of Clinical Laboratory Physicians and Scientists (ACLPS) Jonathan Genzen, MD, PhD University of Utah / ARUP Laboratories Richard Torres, MD Yale University / VA Connecticut Healthcare
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Session 7102
You Want to Measure
“That” in “This”? Body Fluid Testing in the Clinical Laboratory
Academy of Clinical Laboratory Physicians
and Scientists (ACLPS)
Jonathan Genzen, MD, PhDUniversity of Utah / ARUP Laboratories
Richard Torres, MDYale University / VA Connecticut Healthcare
In the past 12 months, neither presenters have had any significant
financial interest or other relationship with the manufacturer(s) of
the product(s) or provider(s) of the service(s) that will be discussed
in our presentations.
Speaker Disclosures
Background & Regulatory Introduction
SPEAKERS AND OVERVIEW
Body Fluid Testing in Clinical Chemistry
Body Fluid Testing in Hematology
Jonathan Genzen, MD, PhDMedical Director, ARUP Automated Core Laboratory
Richard Torres, MDDirector, Yale Flow Cytometry Laboratory
(focus on validations)
PERSPECTIVE
Presenters are NOT:• On the CAP Checklist Committee
• On the CLSI C49 Body Fluid Committee
• Employees of, or speaking on behalf of, CAP, COLA,
CMS, or the FDA
Presenters ARE:• Presenting opinions citing existing regulations and practice
• Presenting ideas for possible improvements
• Providing references to the literature where available
For clarification of regulatory issues,
please contact your accreditation agency
BODY FLUIDSDefinition: “Body fluid, bodily fluids or biofluids are liquids originating
from inside the bodies of living people. They include fluids that are excreted or
secreted from the body as well as body water that normally is not.” (WIKIPEDIA)
All body fluid specimens / spiking sera measured BEFORE mixing (need baseline values)
Measured SPIKED CONC. 1 243 480 760 998
EXPECTED FINAL CONC. 0.9 250.9 500.9 750.9 1000.9([X]fluid x 0.9) + ([X]spike x 0.1)
Calculate PERCENT RECOVERY[X]spiked conc.
expected final conc.X 100
111% 96.9% 95.8% 101.2% 99.7%
[X]fluid
[X]spike
SPIKED RECOVERY
Based on Block and Franke, “Quick Guide to Body Fluid Testing”, AACC Press, 2015.
GOALS: 1) Evaluate ACCURACY
2) Exclude MATRIX EFFECTSPIKED RECOVERY
MIXED RECOVERY
Low Conc High Conc(low end AMR) (high end AMR)
1) Obtain 2 body fluid specimens (low and high)
2) Create a 3rd (50:50 mix)
3) Create 4th and 5th 50:50 mixes (intermediates)
A E
Mid ConcC
Mid-Low ConcB
Mid-High ConcD
100% 75% 50% 25% 0%% of Low0% 25% 50% 75% 100%% of High
Expected = (1.00 x A) + (0 x E) = (.75 x A) + (.25 x E) = (. 5 x A) + (.5 x E) = (. 25 x A) + (.75 x E) = (0 x A) + (1.0 x E)
Measured 1 247 504 734 998
= (1.00 x 1) + (0 x 998) = (.75 x 1) + (.25 x 998) = (.5 x 1) + (.5 x 998) = (.25 x 1) + (.75 x 998) = (0 x 1) + (1.0 x 998)= 1 = 250.25 = 499.5 = 748.75 = 998
PERCENT
RECOVERYMeasured
ExpectedX 100
100% 98.7% 101% 98% 100%
GOALS: 1) Evaluate ACCURACY
2) Exclude MATRIX EFFECT
3) Assess LINEARITY / RANGE
Based on Block and Franke, “Quick Guide to Body Fluid Testing”, AACC Press, 2015.
MIXED RECOVERYGOALS: 1) Evaluate ACCURACY
2) Exclude MATRIX EFFECT
3) Assess LINEARITY / RANGE
DILUTION RECOVERY
High Conc(high end AMR)
A
100% 50% 25% 12.5% 6.25%% of A
Expected
PERCENT
RECOVERYMeasured
ExpectedX 100
100% 95.2% 97.8% 102.6% 99.4%
B C D E1:2 1:4 1:8 1:16
Serial Dilutions
Measured 998 475 244 128 59
Choose high conc. fluid specimen and appropriate diluent
Absolute 998 499 249.5 124.8 62.4
GOALS: 1) Evaluate ACCURACY
2) Exclude MATRIX EFFECT
3) Assess LINEARITY/RANGE
Based on Block and Franke, “Quick Guide to Body Fluid Testing”, AACC Press, 2015.
DILUTION AND MIXED RECOVERYLinearity / Reportable Range
Plot data from Mixing Recovery and/or Dilution Recovery experiments
PRECISIONGOALS: 1) Evaluate IMPRECISION
High Conc. Low Conc.
Run Multiple Times(Within and/or Between Day)
Obtain high and low specimens (or pools)
Analytic Sensitivity• Replicates of specimens near lower limit of quantification
Interferences• Spiking of relevant endogenous / exogenous substances
Stability• Relevant to your laboratory processes
OTHER STUDIES
Reference Intervals• Can be very challenging
• Many fluids do not exist “normally”
• Chart review!
• Literature!
Based on Block and Franke, “Quick Guide to Body Fluid Testing”, AACC Press, 2015.
Tumor Markers (Beckman DXI)
EXAMPLE REFERENCES
Body Fluid Validations (Beckman AU)
Matrix Study (Roche cobas 8000)
Owen et al., Clin Biochem. 2015 Sep;48(13-14):911-4.
RECOVERY STUDIES – PLEURAL FLUID
Roche Cobas 8000
Owen et al., Clin Biochem. 2015 Sep;48(13-14):911-4.
RECOVERY STUDIES – PLEURAL FLUID
Roche Cobas 8000
METHODS AND KITS
Owen et al., Clin Biochem. 2015 Sep;48(13-14):911-4.