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LAB2 - WORKFLOW AND EQUIPMENT Flashcards

1
Q

Unwanted Nucleic Acid into the specimen

A

CONTAMINATION

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2
Q

Repeated Amplification of the same target sequence leads to the accumulation of _____
(10^9) in the laboratory environment

A

Amplification products

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3
Q

Amplification products in PCR is called as

A

amplicons - 10^9

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4
Q

Types of Contamination

A

Contamination to the personnel
Contamination to the product
Contamination to the environment

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5
Q

sources of contamination

A

Cross contamination
Amplification product contamination
Laboratory surfaces
Ventilation ducts
Reagent/supplies
Hair, skin, saliva, and lab clothes of personnel

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6
Q

We process ________ samples each day
especially during this pandemic

A

500-1000

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7
Q

___ means to multiply copy of identical
nucleic acids that is used for diagnosis

A

Amplification

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8
Q

do we have amplicons in pre-analytical phase?

A

naur

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9
Q

COVID-19 testing shall only be done in a laboratory.

A

DOH licensed COVID-19 testing

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10
Q

the agency that will give the license to operate

A

DOH

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11
Q

the agency that will give the necessary compliance and guidelines

A

RITM
HFSRB/CHDRLED.

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12
Q

CHDRLED means

A

Center For Health Development - Regulatory, Licensing, and Enforcement Division

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13
Q

HFSRB means

A

Health Facilities and Services Regulatory Bureau

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14
Q

The COVID-19 testing laboratory shall be a separate
unit, with its own designated working room

true or false

A

true

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15
Q

COVID-19 testing laboratories shall have a _____ (Standard Operating Procedures)

A

Manual of Operations

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16
Q

first section in laboratory workstations

A

specimen receiving area

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17
Q

2nd section in laboratory workstations

A

sample inspevction and lysis room

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18
Q

3rd section in laboratory workstations

A

sample extraction toom

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19
Q

4th section in laboratory workstations

A

reagent mix and prep room

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20
Q

5th section in laboratory workstations

A

template addition room

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21
Q

6th section in laboratory workstations

A

PCR room

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22
Q

7th section in laboratory workstations

A

analysis room

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23
Q

a section that will receive the sample

A

sample receiving data such as oral nasal/oro phareangeal swabs

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24
Q

all samples that would be received by the molecular biology is in __ package

A

triple package

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25
criteria for the triple package
-covered in absorbent material - with resealable plastic and hazard symbol - the resealable plastic must be further placed in another resealable plastic with another hazard p[lus patient identifier
26
who are qualified being receptionist
-any who are graduated in health related course
27
they received samples and ensure that the samples are triple pack and in proper storage such as in ice
receptionisit
28
the section wherein we conduct viral inactivation, and labeling of sample
sample inspection and lysis room
29
how do we conduct inactivation in section 2?
chemical - acid, bases, and those found in VTM physical - temperature (50*C to 60*C)
30
person assigned in section 2 - sample inspection and lysis room
laboratory technician
31
section for obtaining very pure form of nucleic acids
section3 - sample extraction room
32
a section where we use different chemicals to extract nucleic acid
section3 - extraction room
33
an example of chemical which we use in section 3 for nucleic acid extraction
ELUATE
34
in reagent mix and prep room personnel
medtects
35
Reagent used in RT-PCR is _____ which are mixed together
master mix
36
pre-pcr areas or pre analytical area
1. Specimen receiving area 2. Sample inspection & lysis room 3. Sample extraction room 4. Reagent mix preparation room 5. Template additional room
37
called as clean areas due to lack of presence of amplicons
1. Specimen receiving area 2. Sample inspection & lysis room 3. Sample extraction room 4. Reagent mix preparation room 5. Template additional room
38
analytical area
pcr room
39
post analytical area
analysis room
40
LABORATORY WORKSTATIONS MINIMUM PHYSICAL SEPARATION (DOH)
AREA 1- specimen reception (sec 1) AREA 2 - viral inactivation and nucleic acid extraction (section 2,3,5) AREA 3 - reagent storage and handling (section 4) AREA 4 - pcr (section 6) AREA 5 - clerical activities (section 7)
41
out of the area based on minimum physical separation that must be the cleanest
area 3 - section 4 - reagent storage and handling
42
procedures done in section 1
(1) receives and sorts samples; (2) inspects documents; (3) coordinates with healthcare facilities; (4) releases results; (5) facilitates transfer of materials from one station to another; (6) coordinates supplies with supervisor
43
physical set up in section 1
(1) computer; (2) barcode printer; (3) pass box; (4) donning/doffing area; (5) sink
44
ppe in section 1
scrub suits, disposable head cap, surgical mask, nitrile gloves, disposable lab gown, scrub suit, lab shoes or shoe cover
45
section 2 procedures
P7a – inspects and labels with barcode samples inside the BSC; P7b – aliquot samples into two tubes (biobanking and processing); inactivates sample in one tube inside the BSC; P8 – assists & monitors P7a/7b
46
set up of section 2
(1) Class II biosafety cabinets (BSC); (2) heat block inside the hood; (3) centrifuge (4) vortex mixer inside the hood; (5) pipettors & tips; (6) lysis reagent; (7) pass box; (8) refrigerator & freezer; (9) ultra low freezer; (10) biohazard disposal & cleaning supplies & autoclave; (11) labelling area with barcode printer; (12) donning/doffing area; (13) wash area
47
ppe in section 2
scrub suits, lab shoes or shoe cover, lab coat, disposable lab gown, double layer nitrile gloves, , disposable head cap N95/PAPR, face shield
48
procedure in section 3
: P2 – extracts RNA inside the BSC; P3 – assists & monitors P2
49
set up in section 3
(1) class II BSC; (2) heat block; (3) centrifuge; (4) vortex mixer inside the hood; (5) pipettors & tips; (6) extraction kits & ethanol; (7) biohazard disposal & cleaning supplies; (8) refrigerator & freeze
50
ppe in section 3
scrub suits, lab shoes or shoe cover, lab coat, , disposable lab gown, double layer nitrile gloves, disposable head cap surgical mask
51
procedure in section 4
all performed inside the laminar flow hood; (1) prepares mastermix ; (2) dispenses mastermix to PCR plate or tubes; (3) adds negative controls to reaction
52
set up in section 4
: (1) at least laminar flow hood/PCR cabinet (with UV lamp); (2) vortex mixer inside/outside the hood; (3) mini-centrifuge inside/outside the hood; (4) pipettors & tips; (5) PCR reagents; (6) cold rack; (7) refrigerator & freezer; (8) disposal & cleaning supplies
53
ppe in section 4
: scrub suits, surgical mask, nitrile gloves, lab coat, scrub suit, lab shoes or shoe cover
54
procedure in section 6 and 7
facilitates PCR run & result interpretation
55
physical set up in section 6 and 7
: (1) real time PCR machine; (2) computer with software; (3) cold rack; (4) PCR plate or tube centrifuge; (5) disposal and cleaning supplies
56
ppe in section 6 and 7
: scrub suit, surgical mask, nitrile gloves, lab shoes or shoe cover
57
lab equipment biosafety cabinet class required
class II - 70% exhau sted, 30% recirculated
58
autoclaving steam, time, and pressure
15 psi 15 mns 121*C
59
VTM done in processing is stored in
-70*C biobase freezer
60
biobanking stores in what freezer
-70*C biobase freezer
61
frozen bacterial culture
-70*C freezer
62
stores, annotate biological entities
biobanking
63
0.2 ml to 0.5 ml of microcentrifuge tubes is for
pcr
64
1.5 ml tube is for
extraction of nucleic acids
65
for mixing of agarose gel
Spatula
66
solution/components of VTM
hank's balance salt solution fetal bovine serum amphotericin B 400 ug/ml gentamicin sulfate 80 mg/2ml sterile water for injection
67
Hanks; Balanced Salt Solution components
calcium, magnesium, and phenol red, sterile
68
Hanks; Balanced Salt Solution indicator
phenol red
69
used to ensure enought nutrient for VTM
fetal bovine serum
70
antibiotic that inhibits the gowth of fungi in VTM
amphotericin B
71
antibiotic that inhibits the growth of bacteria in VTM
gentamicin sulfate
72
buffer for electrophoresis
tris base
73
act as enhancer of actviity of enzymes
mgcl2
74
biological grade water
nuclease-free water
75
dye for bands of electrophoresis
gel loading dye or buffer
76
gel loading used that is NOT cancerous
GEL RED
77
GEL RED replaced __
ethidium bromide
78
component of PCR kit
dNTPs, MgCl2, Taq polymerase, buffer
79
disrupts protein
Proteinase K
80
reagents stored in room temp
detergents ethanol buffer concentrated solution acids and bases
81
4*C storage
bacterial cultures buffers dna media pcr products serum
82
-20*C storage
enzyme RNA
83
-70*C
frozen bacterial cultures lipids
84
Hazardous Chemicals: we must refer to ______ which describes the composition and properties, toxicology, and instructions for handling, spill control, and waste disposal.
MSDS (Materials Safety Data Sheet)
85
brand for Extraction only
(KingFisher)
86
brand for Extraction & dispensing
(Sansure Natch CS)
87
brand for Extraction, dispensing, and qPCR
bd max
88
the section that is most prominnet in acquiring infection
section2
89

MODX (17 decks)

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