Lab 11: Virus Detection & Transmission MCB 151
LNA: Virus Detection & Transmission
(3 points) Format
LNA must be typed. LNA should contain the headings listed below, with the appropriate
questions/items numbered beneath each heading if indicated. The template provided
for the lab has been utilized.
(5 points) Procedure
Include a detailed description of the process that was performed in Lab 11. Remember
to use your own words. You should provide sufficient detail so someone could repeat the
experiment exactly the way it was performed.
Lab 11's purpose was to detect, examine, and grasp the pandemic of Extrusive Bowel
Syndrome (EBS), a norovirus-related illness. Participants took on three unique roles:
emergency room physician, medical lab scientist, and epidemiologist. Each job
mimicked real-world methods for identifying and managing infectious disease outbreaks.
The first step involved playing the roles of emergency room doctors tasked with
diagnosing two fictitious patients. Using the clinical profiles and case facts provided,
participants investigated the patients' symptoms, which included non-bloody diarrhea,
vomiting, and stomach pain. After determining that the symptoms were consistent with
EBS, the participants collected simulated stool samples for additional analysis. This
initial step ensured that the appropriate samples were transported to the lab for
analysis.
The second step comprised medical lab scientists detecting the presence of norovirus in
stool samples using reverse transcription polymerase chain reaction (RT-PCR).
Participants began by collecting RNA from feces samples in accordance with established
safety standards. RNA was isolated and reverse-transcribed into complementary DNA
(cDNA) using a reverse transcriptase enzyme. Next, specialized primers targeting the
EBS-NoV genome were introduced, and the cDNA was amplified using PCR. Following
amplification, the products were examined using gel electrophoresis. The gel was filled
with PCR products and exposed to an electric current, which separated DNA fragments
based on size. A positive result was revealed by unique DNA bands matching to the virus's
target area, which confirmed the presence of EBS-NoV.
Epidemiological study was carried out in the last phase. Participants used data from the
diagnostic tests and patient histories to track the outbreak's origin and transmission
patterns. A map of the fictional restaurant where the outbreak occurred was supplied. In
order to identify potential locations for viral transmission, participants observed the
movements and interactions of both consumers and staff. With this information, they
were able to construct a chain of infection that included the susceptible host, the
transmission pathway, the entry portal, the reservoir, and the exit portal. This
comprehensive approach demonstrated how to control and halt infectious disease
outbreaks using field, lab, and clinical data.
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Lab 11: Virus Detection & Transmission MCB 151
(25 points) Data Presentation/Results
(3 points) Insert your gel image and label the bands of the MWR using the sizes shown in
Figure 11.8.
Figure 11.8. Agarose gel electrophoresis of the controls and patient samples from the EBS-
NoV diagnostic RT-PCR test. This gel shows the bands obtained including primers that amplify
the EBS-Nov VP1 gene and the human GAPDH gene. MWR = molecular weight ruler; (-)
control = control sample containing the human GAPDH gene; (+) control = positive control
sample contain both the VP1 and GAPDH gene; NTC = no template control (containing no
template DNA); A = sample from Patient A; B = sample from patient B
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Lab 11: Virus Detection & Transmission MCB 151
(8 points) State the results of the control samples and describe what you can conclude
from them.
The results of the control samples show that B7 is positive, indicating an infected
person, but C6 and C3 are negative, demonstrating the absence of illness. E6's status is
unknown and will require more testing to ascertain. Although further research is
necessary for equivocal results, the testing strategy seems to be generally dependable
for identifying positive and negative occurrences.
(4 points, each data point 0.625) Record your restaurant staff/patron results below.
Person ID Infection Status (Positive/Negative/Undetermined)
C6 Negative
C3 Negative
B7 Positive infected
E6 Unknown
(10 points, each data point 0.30) Gather the results from other groups in your section.
In the table below, record the infection status for all who were in the restaurant.
Person ID Infection Status Person ID Infection Status
A1 Negative C6 Undetermined
A2 Negative C7 Positive infected
A3 Positive infected E1 Negative
A4 Negative E2 Negative
B1 Negative E3 Negative
B2 Positive infected E4 Positive infected
B3 Positive infected E5 Undetermined
B4 Negative F1 Negative
B5 Negative F2 Negative
B6 Negative F3 Negative
B7 Positive infected F4 Negative
B8 Negative F5 Positive infected
C1 Negative F6 Positive infected
C2 Undetermined W1 Undetermined
C3 Undetermined W2 Undetermined
C4 Positive infected K1 Positive infected
C5 Undetermined
(38 points) Conclusions
1. (2 points) Droplet and aerosol transmission both involve viruses (or other
pathogens) that are suspended in bodily fluids. Why, then, is droplet
transmission considered direct and aerosol transmission considered indirect?
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Lab 11: Virus Detection & Transmission MCB 151
Droplet transmission is considered direct because illnesses are immediately
transmitted from an infected person to a susceptible host via large respiratory
droplets. The infected individual must be close to the host for these droplets, which
are emitted when coughing, sneezing, or talking, to travel short distances. Aerosol
transmission, on the other hand, is classified as indirect because it contains lighter,
smaller particles with a longer airborne half-life. The transmission method is slower
since these aerosols can spread over greater distances and infect persons who are not
in close contact with the sick person.
2. (2 points) Enveloped viruses have exterior lipid membranes that can dry out in
the environment. Which of the modes of transmission listed in Figure 11.10 is
least likely for enveloped viruses, and why?
Figure 11.10 shows that enveloped viruses are least likely to spread through fomite
transmission (contact with contaminated surfaces or objects). The lipid envelope that
surrounds these viruses is susceptible to environmental conditions and can dry up or
degrade on surfaces, lowering the virus's infectivity. Because of the fragility of the
envelope, these viruses are less persistent in the environment and are more
successfully transmitted via droplets or direct contact, where the envelope remains
intact.
Creating a Transmission Model (34 points)
3. (8 points) On the restaurant diagram (insert diagram in your report):
a. Mark which restaurant staff and patrons were positive based on your
section’s results.
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Lab 11: Virus Detection & Transmission MCB 151
b. Add other information from the patient cards that you think might be
relevant. What did they eat? Did they move about the restaurant?
For example, Person A3, who tested positive for EBS-NoV, ate at the buffet, which
could have been a source of contamination. If others, such as Person B2, ate from the
same part, they may have gotten the virus through contaminated surfaces, utensils,
or the food itself. Furthermore, some people roamed throughout the restaurant,
which could have aided in the spread of the virus, particularly if they interacted with
shared surfaces or came into close contact with others. Person A2, for example, may
have used the restroom after eating and thus encounter possibly contaminated
surfaces. Understanding these motions and food consumption patterns could assist in
determining which regions of the restaurant are most likely to contribute to
transmission.
4. (5 points) Propose a mode and route of transmission for EBS-NoV at the
restaurant. Closely examine the restaurant layout, seating chart, patient
movement, and other available data. Remember EBS-NoV is a novel virus and
may have a different mode of transmission from other NoVs.
EBS-NoV appears to spread mostly via indirect contact with aerosolized particles.
According to the data, the method of transmission might be contaminated surfaces
(fomites) or airborne spread in the restaurant. Individuals who moved throughout the
restaurant, such as Person A3 and E4, most likely spread the infection by touching
shared surfaces. The virus might have been aerosolized by an infected individual
vomiting or coughing, contaminating nearby areas and food outlets.
5. (8 points) Explain how the evidence supports your proposal.
The evidence supports both indirect contact and aerosolized transmission. Several
infected individuals, including A3 and B2, walked around the restaurant, spreading
surface contamination. K1's sickness suggests airborne transmission, as they were
sitting far apart and yet contracted the infection. The concentration of infections
near social areas, such as the buffet and restroom, implies that fomites are a crucial
conduit. Furthermore, individuals such as W1 dealt with several consumers, which
might have assisted spread.
6. (8 points) Build a model to propose how EBS-Nov may have spread through the
restaurant. Include as many of the six points of the chain of infection in your
model as you can.
The infectious agent was the EBS-NoV virus, which was most likely found in sick
patrons or food. Infected persons, workers, or customers who came into touch with
tainted food or surfaces served as the virus's reservoir. Body fluids, such as saliva or
vomit, would have served as the exit point, contaminating surfaces or utensils. The
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Lab 11: Virus Detection & Transmission MCB 151
mechanism of transmission might have been direct contact with contaminated
surfaces, food, or aerosol transfer from sick people. The virus entered the body by
the consumption of infected food or by touching the mouth, nose, or eyes after
encountering contaminated surfaces. Finally, vulnerable hosts included those who had
no immunity to EBS-NoV and came into touch with contaminated locations, such as
other diners and staff.
a. Can you confidently pinpoint the source(s) of the infection? If yes,
who/what was it and how do you know? If no, what other information
would you need?
Without more information, I cannot reliably identify the source of the illnesses. To
find the source, I'd need information on who ate the same items, shared seating
areas, or interacted with the same service workers, as well as any potential
environmental contamination like filthy surfaces or utensils.
7. (3 points) Considering your model, do any of the positive or negative test
results surprise you? How can you explain those results?
I am surprised by test results when those who were near ill people tested negative.
For instance, while sitting adjacent to several sick people, including A3 and B2,
Person A1 tested negative. Variations in viral transmission, such as differences in
immunity or exposure duration, might account for this. It's also possible that certain
people were less likely to get or transmit the virus because they were asymptomatic
or had a lower viral load. The danger of infection may also be increased by
differences in the restaurant's ventilation or cleanliness.
8. (2 points) How might you test your model of how the virus spread?
To evaluate the viral spreading model, the restaurant's design and seating layout must
be thoroughly examined. This involves monitoring people's movements, calculating
their closeness to sick people, and identifying typical exposure areas like bathrooms,
kitchens, and high-touch surfaces. To uncover any hidden exposure patterns, it would
be good to do follow-up testing or contact tracing on people who tested negative but
lived near sick individuals. Laboratory testing may also be undertaken to determine
the presence of viral infection on surfaces such as communal tables, menus, and
utensils. Air quality testing to investigate aerosol transmission would offer further
information about the distribution and possible hazards of sickness.
