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HLA > Laboratory > Flashcards

Laboratory Flashcards

1
Q

Side scatter detects _______ light

A

refracted

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2
Q

Forward scatter detects _______ light

A

diffracted

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3
Q

FITC emits green light at _______ nm

A

525

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4
Q

PE emits orange light at _____ nm

A

575

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5
Q

PerCP emits red light at _____ nm

A

677

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6
Q

APC emits red light at _______ nm

A

660

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7
Q

Photo Multiplier Tubes (PMT) detect

A

Side scatter and fluorescence

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8
Q

Photodiodes detect

A

Forward scatter

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9
Q

Optical filter that transmits wavelengths above a specified cut-off

A

long pass

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10
Q

Optical filter that transmits wavelengths below a specified cut-off

A

short pass

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11
Q

Optical filter that transmits wavelengths in a narrow range around a specified wavelength

A

band pass

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12
Q

FL1 channel measures light emitted from this fluorophore

A

FITC (530nm)

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13
Q

FL2 channel measures light emitted from this fluorophore

A

PE (585nm)

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14
Q

FL3 channel measures light emitted from this fluorophore

A

PerCP (670nm)

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15
Q

Channel values appear linear but are actually _______ values; and the relationship between channels is _______, not additive

A

log; proportional

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16
Q

Channels are used for binning _______ values

A

linear

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17
Q

MESF bead calibration converts channel values to ______ scale

A

linear

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18
Q

Delta MCF increases with ______ incubation temp; ______ serum volume, and ________ cell number

A

increased; increased; decreased

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19
Q

Ficoll isolation cell purity

A

25-80%

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20
Q

Negative selection by Easy Sep cell purity

A

91 - 98%

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21
Q

6 elements of competancy

A
  1. Direct observation of test performance
  2. Recording and Reporting
  3. Record review
  4. Direct observation of maintenance
  5. Test Performance
  6. Problem solving skills
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22
Q

Break even point formula

A

BEP= Fixed costs/(Test price - Variable costs)

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23
Q

Relative Risk formula

A

[a/(a+b)] / [c/(c+d)]

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24
Q

Odds ratio

A

ad/bc

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25
Q

sensitivity

A

a/(a+c)

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26
Q

specificity

A

d/(d+b)

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27
Q

SSO will use _______ conditions for hybridization and _____ conditions for washing

A

non-stringent/stringent

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28
Q

42 CFR part 493

A

CLIA

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29
Q

45 CFR part 160

A

HIPPA

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30
Q

29 CFT part 1910

A

OSHA

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31
Q

Certificate of accreditation is good how long?

A

2 years

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32
Q

CMS can perform random inspections but will give lab _____ advanced notice

A

2 weeks

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33
Q

Changes in name, ownership, location or director of lab must be reported to _____ and _______ within _______.

A

CMS/ASHI/30days

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34
Q

Off site archived records must be retrievable within _____

A

48 hours

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35
Q

Competency must be performed ________ during first year and ______ thereafter. Also must be completed with ________ changes. Competency is not required for _______ unless they perform tests.

A

semiannually/annually/instrument or method/director

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36
Q

A new director must review and sign all procedures within ______

A

6 months

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37
Q

Cell control panel used for specificity validation must have 3 controls

A
  1. Positive for antigen
  2. Negative for antigen
  3. Positive for corrective antigen
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38
Q

Computer systems must have assessment performed this often

A

1 year

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39
Q

Calibration performed every _______ and during certain situations (name 3)

A

6 months

  1. Change in reagents
  2. Major maintenance or replacement of parts
  3. Controls are out of limits and other trouble shooting has failed
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40
Q

Lab must verify and document thermal cycler temperature accuracy this often

A

6 months

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41
Q

Continuing education requirement of ARB by ASHI: Director

A

50 hrs/year

42
Q

Continuing education requirement of ARB by ASHI: General supervisor

A

27 hrs/year

43
Q

Continuing education requirement of ARB by ASHI: Tech

A

12 hrs/year

44
Q

Continuing education requirement of ABHI: CHA

A

60 hrs/3 yrs

45
Q

Continuing education requirement of ABHI: CHT

A

60 hrs/3yrs

46
Q

Continuing education requirement of ABHI: CHS

A

80 hrs/3yrs

47
Q

Continuing education requirement of ABHI: Director

A

150 hrs/3yrs

48
Q

Education requirements for Tech (3)

A
  1. Medical doctor or doctoral, masters, or bachelor’s degree in science
  2. Associate degree in lab science or medical technology
  3. 60 semester hours of which 24 hours should be medical laboratory technologist school OR 24 hours of basic science course (of which 6 hrs are chem, 6 hrs are bio, and 12 are miscellaneous science courses)
49
Q

Methionine is the ______ codon

A

start

50
Q

Methionine codon

A

AUG (ATG)

51
Q

Stop codons

A

UGA (TGA), UAA (TAA), UAG (TAG)

52
Q

Flow cytometry: must run _______ and _______ standards each time instrument is ________, _________, ______

A

optical/fluorescent/turned on/maintained/adjusted or fix due to problem

53
Q

Controls must be run this often

A

daily

54
Q

Controls for quantitative assays

A

2 controls of different concentrations

55
Q

Controls for qualitative assays

A

Negative and Positive control

56
Q

New lots: Parallel testing with previous lot must be performed; the number of tests to run in parallel is determined by ________

A

technical supervisor

57
Q

Cell culture: Laminar flow hood must be used for cultures incubating _______

A

> 18 hrs

58
Q

Cell culture: Incubators must be monitor for _____, ______, ______

A

temperature/CO2 concentration/humidity

59
Q

MLC must have these controls

A
  1. Negative control for each responder cell consisting of responder cells stimulated by autologous cells
  2. Positive control for responder cell proliferation consisting of known HLA Class II disparate stimulator cells from unrelated donors as well as pooled donors.
60
Q

Serum used in MCL culture medium should be screened for these 3 things

A
  1. Ability to support cellular proliferation
  2. Lack of cytotoxic antibodies
  3. Sterility
61
Q

SSP must have these controls

A
  1. Internal control to detect technical failure

2. Negative control to detect contamination

62
Q

NGS: ________ must be monitored by rotating control samples with different barcode sequences

A

fidelity

63
Q

All UNOS histocompatibility labs must review and verify UNET data for each patient and documentation of this review must be maintained for at least _______

A

3 yrs

64
Q

STR: If subpopulations are analyzed, ______ of each cell subset must be obtained and documented

A

purity

65
Q

Reports must include at minimum

A
  1. Lab name
  2. Director’s name
  3. Lab address
  4. CLIA/ASHI #
  5. Date of collection
  6. Date of report
  7. Unique patient identifier
  8. Method
  9. Result
66
Q

There must be documentation of condition and disposition of ________ specimens

A

rejected

67
Q

Parallel testing between instruments, sites, and methods must be performed this often

A

q 6 months.

68
Q

Approximate length of Class II amplicon

A

270 bp

69
Q

Approximate length of Class I amplicon

A

1000 bp

70
Q

Requirements for general supervisor (5)

A
  1. lab director
  2. technical supervisor
  3. Medical doctor degree + 3 ys of HLA lab experience
  4. Doctoral, masters, bachelor’s degree in science + 3 yrs of HLA lab experience
  5. Served as General Supervisor before 2/28/92 and functioned as technologist + 6 years of lab experience, 2 of which in HLA lab
71
Q

NFPA Diamond: _____ red square is for ________

A

Top/Fire hazard (flash point - will not burn to <73F)

72
Q

NFPA Diamond: _____ yellow square is for ________

A

Right/Reactivity (volatility - stable through may detonate)

73
Q

NFPA Diamond: _____ white square is for ________

A

Bottom/Specific hazard (e.g., oxidizer, acid, alkali, corrosive, radiation)

74
Q

NFPA Diamond: _____ blue square is for ________

A

Left/Health Hazard (Normal to Deadly)

75
Q

2 methods used in screening for Class II antibodies using cell based assays

A
  1. Parallel screening using B and T cell targets

2. Absorb out Class I antibodies with platelets

76
Q

MLC: Responses considered non-stimulatory if stimulation index is _______

A

<2.5

77
Q

Stimulation index formula

A

(A/Bx)/(A/Ax) [Response to allogeneic stimulator cell/Response to autologous stimulator cell]

78
Q

MLC: Patients with ________ can have unresponsive cells making test uninterpretable

A

leukemia

79
Q

Best statistical method for evaluation of serologic typing reagents

A

correlation coefficient

80
Q

Optimizing _______ can prevent plateau effect in PCR

A

number of cycles

81
Q

CSFE is a _________ dye

A

supravital

82
Q

trypan blue is a _______ dye

A

vital

83
Q

Thawing solutions without mixing can result in _________

A

solute gradients

84
Q

EBV transformed B-cells for HLA testing can result in ________ due to ________.

A

cytotoxicity/false positive; non-HLA antibodies

85
Q

Probes in SSO generally bind membrane distal portion of HLA molecules, which refers to ________

A

top, exposed portion of molecule (i.e., the peptide binding portion, where the polymorphisms lie)

86
Q

HCFA stands for

A

Healthcare Finance Administration

87
Q

Anti-T cell therapy (ATG) can cause _______ on antibody assessment

A

false Class I reactivity

88
Q

__________ areas should be physically separated in oder to prevent DNA contamination

A

Pre-amplification

89
Q

AMOS XM introduces __________ prior to complement to prevent ___________ factors

A

wash steps; anticomplementary

90
Q

_________ is considered to be the least sensitive XM method

A

Standard NIH

91
Q

Selection of sera for XM of allosensitized patients should be based on ________ and ___________

A

historic and current sensitizing events.

92
Q

Medicare Part A covers reimbursement for HLA testing during __________ period

A

pre-transplantation

93
Q

_________ annealing temperature can ________ specificity in allele-specific PCR

A

Increasing/increasing

94
Q

Low control in lymphotoxicity assay can be the result of _______ contamination or _______ control serum

A

T-cell/out-dated

95
Q

MLC: Stimulator cells are irradiated with _______ rads to inhibit ________ but preserve _______

A

3000 - 4000/DNA synthesis/Class II expression

96
Q

MLC: increase reactivity of autologous control can be caused by _______, ________, and ______

A

viral infections, recent transfusions, leukemia relapse

97
Q

One-way primary MLC is basically a ___________

A

cellular XM procedure

98
Q

Methods for HLA antigen isolation/purification

A

detergent solubilization and affinity chromatography

99
Q

MLC: limiting dilution analysis calculates frequency of functional lymphocyte populations based on assumption that those populations follow a _________ distribution

A

Poisson

100
Q

Isolating of peptides from HLA molecules can be performed after isolating the HLA molecule by _______ of peptides

A

acid elution

HLA (11 decks)

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