Specimen Collection Flashcards

(83 cards)

1
Q

_____ is not as critical for most molecular testing. The quality of nucleic acids may be compromised if the specimen is ______ handled.

A

Viability

improperly

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2
Q

DNA and especially RNA will be damaged in ____ or ______. Due to the ____________, it is also important to __________ that could yield false positive results.

A

lysed or nonviable cells
sensitivity of molecular testing
avoid contamination

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3
Q

Sampling must include material from the ______infection. The time and site of collection should be _____for the likely presence of the infectious agent.

A

original

optimal

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4
Q

The quantity of target organisms, as well as clinical implications, should be taken into account when interpreting the significance of _________.

Molecular detection can reveal infective agents at levels ______ clinical significance Conversely, _____________ methods may miss detection of a variant organism.

A

positive results
below

highly specific molecular

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5
Q

Blood draws should go into the proper _________, fi one is to be used.

A

anticoagulant

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6
Q

________ swabs may be used for throat cultures.
___________swabs with plastic shafts have been recommended for collection of bacteria, viruses, and mycoplasma from mucosal surfaces.

A

Wooden-shafted swabs

Dacron or calcium alginate

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7
Q

_______are less adherent to the microorganisms and will not interfere with PCR reagents, with the exception of calcium alginate swabs with __________, which had been reported to affect PCR amplification.

A

Plastics

aluminum shafts

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8
Q

Collection methods such as____________, ______,_______ have been designed for maximum recovery of microorganisms from swabs by centrifugation.

o _________– sterile cups, screw-capped tubes, stoppered tubes, petri dishes
o ________ – calcium alginate swabs, Dacron swabs, cotton swabs, nasopharyngeal-urogenital swabs, swab transport system
o ________ – Neisseria gonorrhoeae transport systems, swab extraction tube system
o ___________ – molecular testing, N. gonorrhoeae transport systems, STAR buffer
o ___________s – Starplex anaerobic transport system (Fisher), BBL vacutainer anaerobic specimen collector
o _________ – BD cellmatics viral transport pack, BBL viral culturette

A

swab extraction tube system. sonication, and vortex

Sterile containers 
Swabs
Specialty systems
Proprietary systems
Anaerobic transport system
Viral transport systems
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9
Q

Isolating nucleic acids from microorganisms is similar to
isolating nucleic acids from _______ with only a few
additional considerations

A

human cells

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10
Q

Isolating nucleic acids from microorganisms is similar to isolating nucleic acids from human cells with only a few additional considerations.

o Depending on the microorganism, more________ procedures may be required. Mycobacteria and fungi, in particular, have ____ cell walls that are more difficult to lyse than those of other bacteria and parasites. __________ has a thicker cell wall than gram-negative bacteria and may require more rigorous cell lysis conditions. ______ lacks a cell wall, and so care must be taken with the sample to _________of the cells and loss of nucleic acids.

A
rigorous lysis
thick
Gram-positive bacteria
Mycoplasma
avoid spontaneous lysis
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11
Q

The concentration of organisms within the clinical sample must be considered. Samples can be ________ to concentrate the organisms within the fluid from the __________ that are often received down to volumes that are appropriate for molecular procedures.

A

centrifuged

milliliters of sample

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12
Q

Inhibitors of enzymes used in molecular analysis may
be present in __________; removal or
inactivation of inhibitors might be included in
_________ methods.

A

clinical specimens

specimen preparation

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13
Q

If RNA is to be analyzed, _________of
RNases in the sample and in all reagents and
materials that come into contact with the sample is
important.

A

inactivation or removal

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14
Q

When processing a whole blood specimen, it is
important to remove _______ and other
products of __________because they
can ___________ and thus may ___________ of nucleic acid in the sample, resulting in a _________ PCR result

A
hemoglobin
metabolized hemoglobin 
inhibit DNA polymerase
prevent 
the amplification
false negative
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15
Q

In blood samples, white blood cells are isolated from
the red blood cells using _________ and then
lysed.

A

Ficoll-Hypaque

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16
Q

Whole blood is processed in _____________, which effectively __________ and other contaminating molecules.

A

automated DNA isolation systems

remove hemoglobin

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17
Q

__________, devoid of red blood cells, are also

used as sources of microorganism nucleic acid.

A

Serum and plasma

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18
Q

Sputum is a source of nucleic acid from organisms

that cause ______________. __________ present in sputum may inhibit DNA polymerase and thus must be removed.

A

respiratory tract infections

Acidic polysaccharide

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19
Q

___________ is treated similarly to cerebrospinal fluid; that is, the specimen is _____________the organisms and then subjected to nucleic acid isolation procedures. Inhibitors of DNA polymerase -_________________________ – have been demonstrated in urine as well.

A

Urine
centrifuged to concentrate
nitrate, crystals, hemoglobin, and beta-hCG

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20
Q

Any clinical specimen _____ used as a source of

microorganism nucleic acid for analysis.

A

can be

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21
Q

Depending on the specimen,_______________ may be necessary to allow for optimal nucleic
acid isolation, amplification, and analysis

A

special preparation procedures

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22
Q

The presence of inhibitors of DNA polymerase has been demonstrated in clinical samples.

Careful separation of nucleic acid from other molecules present in the sample will _____________.

The type of specimen used for molecular testing will also affect extraction and yield of nucleic acid.

Reagents and devices have been developed to combine collection and extraction of nucleic acid from difficult specimens.

A

ensure target amplification

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23
Q

The presence of inhibitors of DNA polymerase has been
demonstrated in clinical samples.

Careful separation of nucleic acid from other molecules
present in the sample will _____________.

The type of specimen used for molecular testing will also
affect extraction and yield of nucleic acid.

Reagents and devices have been developed to combine
collection and extraction of nucleic acid from _________

A

ensure target amplification

difficult specimens.

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24
Q

Molecular tests require _______specimen handling and processing for ______ and_____ test results. The success of a test procedure is affected by the ____,_____and ______ of specimens.

A

optimal
accurate; consistent
age, type, condition

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25
___________– consequence of erroneous or misleading results caused by events that occur prior to sample analysis.
Preanalytical error
26
Each laboratory will have requirements for specimen handling, but __________ apply to all specimen collection. The condition of the specimen and the chain of custody is reviewed on__________in the laboratory
general policies | receipt
27
If a specimen shows evidence of tampering or is hemolyzed, degraded, clotted or otherwise compromised, the technologist must ____________.
notify the superviso
28
No specimen is accepted___________ and identification on the specimen tube or container, nor is a specimen accepted if the labeling on the ______________ that on the accompanying requisition.
without proper labeling | specimen does not match
29
If a specimen is unacceptable, the ____________ of the specimen is recorded in the _________ or laboratory quality assurance records.
disposal or retention | patient report
30
If not processed immediately, specimens are maintained in ________________under the appropriate conditions for the analyte being tested.
secure areas with limited access
31
For routine specimens tested by ___________, the entire specimen is often not used. If more than one test is to be performed on the same specimen, ___________ of specimens is carefully avoided. This can most likely occur from ________ carryover.
amplification methods cross-contamination pipetting
32
An aliquot removed from a specimen is __________ to the original tube or vessel. Molecular genetic tests may require dedicated specimens not shared with other tests.
never returned
33
________ inhibits enzyme activity. Specimens should be inspected for visual signs of _________. ________ and _______ are removed effectively in most DNA and RNA isolation procedures; however, if white blood cell lysis has also occurred, DNA or RNA yield will be reduced. This could result in _______ results in qualitative testing or _______________in quantitative analyses.
``` Hemoglobin hemolysis Hemoglobin and coagulants false-negative inaccurate measurements ```
34
______ and _________amplification are routinely performed on paraffin-embedded tissue samples. ______________, requiring large fragments of DNA, are less likely to work consistently with fixed tissues.
PCR and RT-PCR | Southern or northern blot
35
Collection Tubes for Molecular Testing: ``` 1. ____________ – chemistry, serum, viral antibody studies 2. ______________ – immunology, virology studies 3. ____________ – cytogenetic studies, molecular studies 4. ____________ – virology, molecular biology studies 5. ______________ – molecular biology studies ```
``` (Red) No additive (Green) Sodium heparin freeze dried (Brown) Sodium heparin (Lavender) Tripotassium EDTA (Yellow) Acid citrate dextrose solution ```
36
___________– recommended by the CDC for handling potentially infectious specimens. All specimens are potentially infectious, so they should all be handled with standard precautions using proper personal protective equipment to prevent disease transmission. Standard precautions, including ______ as PPE, are used by the molecular laboratory technologist who has no direct contact with patients.
Standard precautions
37
____________ precautions – include respirators; used with airborne or contact-transmissible agents.
Transmission-based
38
_______________ – for direct patient care where there is the potential for direct exposure to infectious agents on or form the patient.
Contact precautions
39
_________ are also important to protect nucleic acids from nuclease degradation and are absolutely required for handling of RNA.
Gloves
40
_____ is less susceptible to degradation from contaminating | DNases.
DNA
41
Repeated handling of samples _________ will | adversely affect the integrity of the DNA over time.
without gloves
42
Standards and controls that are handled _________ are the most likely to be affected. Having separate areas for DNA and RNA isolation is ______.
repeatedly | recommended
43
Change in assay response with corresponding change in analyte. Ø Ex: All positive reference standards tested positive with the new assay. The analytical sensitivity of the assay is 100%.
Analytic sensitivity
44
Ø Ability of test result to predict a clinical condition. Ex: Of 100 patients with a gene mutation, 95 have a disease state, a clinical sensitivity of 95%.
Clinical sensitivity
45
Ø Least detectable presence of the analyte Ex: The t(14;18) translocation test can detect 1 translocated cell in 10, 000 normal cells, a detection limit of 0.01%.
Detection limit or Limit of detection
46
Ø Ability to detect only the analyte and not nonspecific targets Ex: The invader assay for factor V Leiden successfully detected mutations in 18 positive specimens while yielding negative results for 30 normal specimens; no false positives.
Analytic specificity
47
Ø Disease-associated results only in patients who actually have the disease conditions Ex: Of 100 normal specimens, 1 displayed a gene mutation; 1 false positive, a clinical specificity of 99%.
Clinical specificity
48
Ø Agreement between independent test results. Ø Ex: A quantitative method yields 99 results less than 1 standard deviation apart in 100 runs, a precision of 99%.
Precision
49
Ø Consistency of test results produced from the same procedure. Ø Ex: A qualitative method yields 100 positive results when performed in 10 independent laboratories, a reproducibility of 100%.
Reproducibility
50
Ø The range within which a specimen may be measured | directly without dilution or concentration.
Analyte measurement range
51
Ø The range within which test results are considered to | be valid with or without dilution.
Reportable range
52
Ø Expected analyte frequency or levels from a population of individuals. Ø Ex: The reference range for prostatic specific antigen is 0-4 ng/mL.
Reference range
53
Ø Production of correct results. Ø Ex: Of 100 specimens with mutations in the HCM gene, 99 are detected by sequencing, with no mutations detected in normal specimens.
Analytic accuracy
54
Ø Quantitative correlation between test result and actual amount of analyte. Ø Ex: A graph of test procedure results versus input analyte yields a straight line.
Linearity
55
_________– establishment of conditions for instrument response/method result association with the true value of quantitative analyte measurements.
Calibration
56
__________ – samples of known amounts/concentrations of molecules of the same type measured in the assay, such as an admixture of PCR products in known proportion or an RNA transcript calibrator in a matrix of normal RNA. The source, quality, and preparation of the calibrators should be documented.
Calibrators
57
______________– performed if calibration does not | meet the required standards of linearity or accuracy.
Re-calibration
58
_____________ – may be required upon changes of or major repairs to instruments or changes in reagent lots that might affect test performance or when quality control indicates shifts or unacceptable errors in test control results. Verification should demonstrate the continued linearity of correlation between the calibrator values and test results.
Calibration verification
59
These are samples of known type or amount that are | treated like and run with patient specimens.
controls
60
Interpretation of test results always includes inspection of ______and ______ to verify acceptable test performance.
controls and standards
61
The controls and standard curve should cover the ___________levels or results of the method. __________are continually monitored to spot trends or spikes outside of tolerance limits.
critical detection Control results
62
___________– defines lower limit of detection for more meaningful interpretation of negative results. For qualitative tests, along with positive and negative controls
Sensitivity control
63
_______________– a target should always amplify. It distinguishes true-negative amplification results from false negatives resulting from amplification failure. For nucleic acid amplificationtechniques. In quantitative methods, high-positive, low-positive, and negative controls are included with each run. The high and low levels should be similar to critical points in the assay, such as the lowest detectable level of analyte.
Amplification control
64
____________ – or dilution series of analyte levels encompassing the levels expected from the patient specimens. Real-time PCR methods automatically determine analyte levels that require measurement of this curve or dilution series.
Standard curve
65
____________ – run in the same reaction mix as the test specimen; in methods requiring detection of a target-specific product or relative amounts of target.
Internal control
66
____________________ should be | included in each run of patient samples
Positive, sensitivity, and negative controls
67
For tests with multiple targets, controls can be systematically rotated for different targets in each run. • Controls for quantitative tests should reflect the __________ levels of test results. • _________ or acceptability levels of control values should be established based on the validated precision and reproducibility of the assay.
clinically critical | Tolerance
68
Monitoring can be performed using ___________ or modified LJ plots or cusum plots, and tolerance limits can be expressed as __________. If the control results violate the set limits, then corrective action such as _____________has to be taken and documented.
Levey-Jennings Westgard rules repeating the run with replacement controls
69
If patient specimen availability precludes repeating a run, ___________ into the nature of the ___________ and ___________ result ____ allow acceptance of the sample results without repeating.
further investigation control failure its effect on the patient may
70
Comparison of test values with previous results or historical averages may _______________ in the test results
serve to increase confidence
71
Monitored controls and conditions are reviewed ________ or at determined intervals to document outliers or systemic error.
monthly
72
Periodic review and documentation of test results are | ______ for all clinical testing, including molecular tests
required
73
Critical values that require physician notification are | established by ________ and confirmed by _______.
validation | monitoring
74
Assay levels that distinguish positive from negative results, cut-off values or cut points, must be _______ and verified at regular intervals
well defined
75
must be well defined and verified at regular intervals. Also includes documentation and maintenance of laboratory procedures and methods. These procedures are frequently updated with events ranging from major advances in instrument technology to changes in reagent availability.
76
Test results may show _________ with other laboratory findings, with clinical observations, or with the laboratory’s own preliminary results.
discrepancies
77
____________ are documented along with any corrective | action taken, if necessary.
Discrepancies
78
Due to the nature of molecular pathology testing, discrepancies may be explained by the technical aspect of the test.
o Increased sensitivity o High resolution o Varied methodologies used
79
HAZARDOUS MATERIAL CLASSIFICATION 1 2 3 W
REACTIVITY FIRE HAZARD HEALTH HAZARD SPECIFIC HAZARD
80
SPECIFIC HAZARDS: W
``` OXIDIZER ACID ALKALI CORROSIVE USE NO WATER RADIATION ```
81
REACTIVITY ``` 4 3 2 1 0 ```
``` May deteriorate Shock and heat may deteriorate Violent chemical change Unstable if heated stable ```
82
FIRE HAZARD ``` 4 3 2 1 0 ```
``` Below 73*F Below 100*F Below 200*F Above 200*F Will not burn ```
83
HEALTH HAZARD ``` 4 3 2 1 0 ```
``` Deadly Extreme Danger Hazardous Slightly hazardous Normal material ```