Compilation of Rules and Regulations of the State of Georgia › Department 111 - RULES OF DEPARTMENT OF COMMUNITY HEALTH › Chapter 111-8 - HEALTHCARE FACILITY REGULATION › Subject 111-8-10 - LICENSURE OF CLINICAL LABORATORIES › Rule 111-8-10-.22 - Quality Control for Flow Cytometry

Current through Rules and Regulations filed through June 27, 2024
(1) A standard, consisting of latex beads or
				other uniform particles, shall be run to insure proper focusing and alignment
				of all lenses in the path, and for both the existing light source and signal
				(light scatter, fluorescence, etc.) detectors. The results of optical focusing/
				alignment must be recorded each day.
(2) A threshold value for acceptable optical
				standardization shall be established for all relevant signals for each
				instrument and the focusing procedure repeated until these values are achieved
				or surpassed. If a particular threshold value cannot be attained, a written
				protocol for instituting corrective action must be available and
				used.
(3) A fluorescent standard,
				for each fluorochrome to be used, shall be run to insure adequate amplification
				of the fluorescent signal(s) each day of use and after any maintenance or
				adjustment of the instrument. This standard may be incorporated in the beads or
				other particles used for optical standardization or may be a separate bead of
				fixed cell preparation. If acceptable fluorescence separation cannot be
				attained, a written protocol instituting corrective action must be available
				and used.
(4) When performing
				analyses using two or more fluorochromes simultaneously, an appropriate
				procedure must be used to compensate for "spill over" into the other
				fluorescence detectors.
(5) For
				laser based instruments, the current input (amps) and laser light output
				(milliwatts), at the normal operating wavelength measured after the laser is
				peaked and normal operating power set, must be recorded as part of a daily
				quality control record.
(6) The use
				time of instruments with mercury fluorescent lamps must be recorded. Lamps must
				be replaced when the allowable use limit has been reached.
(7) The laboratory must run a positive and
				negative control each day of patient testing. The negative controls should
				include normal serum from a healthy individual. The positive control (using
				appropriate dilutions) should react with cells representing all HLA types
				(i.e., pulled high PRA sera).
(8)
				Each laboratory must establish and document its own threshold with multiple
				normal sera for discriminating positive crossmatches. For significant change in
				protocol, instrumentation, or software, the characterization of the positive
				threshold must be repeated.
(9) For
				internal labeling, the method used to allow fluorochrome labeled antibodies to
				penetrate the cell membrane must be documented as effective.
(10) Whether analyzed directly or fixed prior
				to analysis, labeled cells must be analyzed within a time period demonstrated
				by the laboratory to avoid significant loss of any cell sub-population or total
				cell numbers. Test samples must be analyzed within the same period, after
				staining, as the control samples.
(11) If analysis will be based on a
				population of cells selected by flow cytometry "gating" on size or density
				parameters, or selected depletion or enrichment techniques, control stains must
				be run for each test individual to detect the presence of contaminating cells
				in the selected population (i.e., monocyte contamination of lymphocytes gated
				by forward angle or forward angle vs. 90 degree light scatter must be detected
				with a monocyte specific marker antibody). For cell surface labeling, a method
				must be used to determine the proportion of viable cells in the population of
				counted cells (i.e., thidium bromide staining, CD45 staining). If this value
				falls below an established laboratory threshold, an appropriate reanalysis of
				specific measurements must be done.
(12) Conclusions about abnormal proportions
				of abnormal cell surface marker shall only be drawn in comparison with local
				control data obtained with the same instrument, reagents and techniques.
				Conclusions about leukemia/lymphoma classification shall be based on local or
				published reference data. Determination of percent positives must take into
				consideration the results of the negative control reagent.
(13) The specificity of monoclonal antibodies
				shall be verified by the published and/or manufacturer's documentation and,
				whenever possible, verified locally through tests with appropriate control
				cells prepared and tested by the same analysis. The quantities of reagents used
				for each test sample must be determined by the manufacturers from published
				data and, whenever possible, should be verified locally by appropriate
				titration procedures.
(14)
				Terminology used must be defined and/or must conform to nomenclature
				recommended/approved by the most recent International Workshop of
				Differentiation Antigens of Human Leukocytes or other appropriate scientific
				organizations.
O.C.G.A. §§
										31-22-1,
				  31-22-4.