M5

Question 1

the gold standard for the detection of SARS-CoV-2 due to the sensitivity and specificity
of the test.

Answer 1

Reverse-transcription PCR (RT-PCR)

Question 2

Two most common tests used to confirm COVVID19:

Answer 2

RTPCR and ELISA

Question 3

powerful diagnostic tool in medicine. This technology rapidly and
reliably amplifies specific sections of DNA

Answer 3

PCR

Question 4

____, allows a diagnostic data that converts RNA to DNA
and then amplify copies of the viral genome that are present.

Answer 4

A modified PCR known as RT-PCR

Question 5

Examples of RNA Viruses

Answer 5

1. Dengue Virus 4. Ebola Virus Disease 7. Measles
2. Hepatitis C and E 5. Rabies 8. COVID-19
3. West Nile Fever 6. Polio

Question 6

Each PCR cycle (D → A → E) ____ the amount of the target DNA in less than ___

Answer 6

doubles, five
minutes

Question 7

___ cycles may be required to produce enough DNA for analysis

Answer 7

20 to 40

Question 8

primers are combined into a single reaction on the PCR

Answer 8

Multiplex reaction

Question 9

Multiplex tests also allow an internal
control to be amplified in every ___single reaction.

Answer 9

single

Question 10

The control is often a common DNA
sequence found in humans, called a ____, that indicates whether or not
the experiment was successful

Answer 10

“housekeeping gene”

Question 11

PCR was invented by ___ in ___

Answer 11

Dr. Kary Mullis in 1984.

Question 12

He recognized that he could replicate DNA in vitro using ____ and ___ in a process similar to DNA replication
in a cell’s nucleus

Answer 12

short, synthetic DNA
oligonucleotides (primers) and DNA Polymerase I

Question 13

____ father of DNA studies; assisted by Rosalind Franklin

Answer 13

Watson and Crick

Question 14

This technology was then supercharged by ___

Answer 14

Dr. Randy Saiki

Question 15

He began using thermostable DNA polymerase from the bacteria ____ in PCR reactions

Answer 15

Thermus aquaticus called
Taq

Question 16

 Use of Taq polymerase in PCR was announced by ___

Answer 16

Henry Erlich

Question 17

The announcement as made in a meeting in___ on ___, submitted for
publication in October 11987, and was published early the next year

Answer 17

Berlin on September 20, 1986

Question 18

The patent for PCR
with Taq polymerase was filed on ___, and was issued on October 23, 1990

Answer 18

June 17, 1987

Question 19

Purified double-stranded DNA is mixed with primers, Taq
polymerase, and nucleotides

Answer 19

DENATURATION:

Question 20

94 c

Answer 20

DENATURATION:

Question 21

Cool sample to 45-60⁰C to allow primers to base pair with the target
DNA sequence

Answer 21

ANNEALING:

Question 22

Temperature is raised to 72⁰C, the optimal temperature at which Taq
polymerase will extend the primer to synthesize a new strand of DNA for analysis

Answer 22

EXTENSION

Question 23

ability to test to correctly identify an
individual with a disease as positive

Answer 23

sensitivity

Question 24

Ability of a test to correctly identify an
individual without the disease as negative

Answer 24

specificity

Question 25

Technique used to analyze amplified DNA

Answer 25

Agarose Gel Electrophoresis

Question 26

The RNA is reverse transcribed into
complementary DNA (cDNA), using __

Answer 26

reverse transcriptase.

Question 27

SARSCOV2 has ___ main structural proteins.

Answer 27

four

Question 28

The single stranded
DNA molecule is then completed by the ______ activity of the
reverse transcriptase into cDNA

Answer 28

DNA-dependent DNA polymerase

Question 29

 Monomers of the nucleocapsid of N protein link together to form a ___ which wraps
around and protects the RNA genome.

Answer 29

helical capsid

Question 30

 Embedded in the membrane are several viral proteins: ____

Answer 30

the spike or S protein, the envelope
or E protein, and the membrane or M protein.

Question 31

The____protein coordinates interactions between the other viral proteins and the
host cell factors, turning cells into virus factories

Answer 31

membrane

Question 32

As a ___, the envelope protein binds to itself to form channels that facilitates viral
release.

Answer 32

viroporin

Question 33

The
____ protein binds with human cell surface protein allowing the virus to inject its genetic
material into its host cell

Answer 33

spike

Question 34

. An intermediate or inconclusive test occurs
when only one of the two SARSCOV targets amplify.

Answer 34

presumptive positive

Question 35

 All buffers except ___ and ____ in this kit can be stored at 15⁰C - 35⁰C

Answer 35

Proteinase K and Carrier molecule

Question 36

Proteinase K should be stored at ____

Answer 36

2⁰C – 4⁰C after reception.

Question 37

Carrier molecule should be stored at P_____. Carrier molecule is
not recommended to be frozen and thawed more than ___

Answer 37

-25⁰C to -15⁰C after dissolution, 8 times

Question 38

in dry sample, Put the dry swab into ___of Swab/Stool lysis buffer and vortex thoroughly.

Answer 38

1 mL

Question 39

Prepare 0.4% DTT solution.

Answer 39

in sputum

Question 40

Put ____of fecal sample into 1.5 ml tube

Answer 40

180-220 mg or 200 uL

Question 41

10 minutes incubation

Answer 41

dry sample
fecal sample treatment

Question 42

In fecal sample, Centrifuge for ____at ___

Answer 42

3 minutes at 13000 rpm 5

Question 43

pre treated sample

Answer 43

200 ul

Question 44

microcentrifuge

Answer 44

1.5 ml

Question 45

ksb buffer

Answer 45

250 ul

Question 46

proteinase k

Answer 46

200 ul

Question 47

carrier molecule

Answer 47

5 ul

Question 48

Vortex thoroughly for ___ to mix and Incubate at___ for

Answer 48

10 seconds, 56⁰C for 10 min

Question 49

Absolute ethanol Vortex

Answer 49

350 uL

Question 50

After dispensing all the mixture on the provided spin column, centrifuge at ___ for __

Answer 50

10,000 rpm for
1 minute.

Question 51

● Unicellular
● Can survive without host
● Have DNA

Answer 51

bacteria

Question 52

Unicellular
● Cannot survive without host
● Only classified as RNA or DNA
● Many viruses don’t have DNA

Answer 52

virus

Question 53

Was awarded Nobel Prize Chemistry in 1993
- Used fragments of E.coli DNA polymerase to
describe the in-vitro amplification of genes

Answer 53

KARY MULLIS, Ph. D

Question 54

First proposed by___

Answer 54

H.G. Khorona, et al (1970s)

Question 55

used in diagnostic PCR

Answer 55

MULTIPLE PRIMERS

Question 56

Base lane
● Intended for analyte of interest

Answer 56

lane 1

Question 57

SARS-COV 1

Answer 57

• 1992-2002

Question 58

MERS-COV

Answer 58

• 2012 peak