Veterinary Epidemiology

ONLINE ACTIVITY
Topic: Evaluation and Interpretation of Diagnostic Tools

NAME: Mendoza, Amber Lee G._______________________________________

Article for Review: https://bmcinfectdis.biomedcentral.com/articles/10.1186/s12879-019-4277-8

Guide Questions:

1. What was the diagnostic platform being studied in the paper? Explain briefly the background of
this diagnostic platform.

Reverse Transcriptional Loop-mediated Isothermal Amplification (RT-LAMP) , combined with

one-pot colorimetric visualization system. Detection is accomplished by visual inspection of
color of the agar for a positive.

2. What are the viruses that should be detected by RT-LAMP? (Enumerate)

Seasonal influenza viruses and Avian influenza viruses

3. To which another diagnostic test was RT-LAMP was compared to in terms of its sensitivity?
conventional RT-PCR-based

4. To determine the specificity of the multiplex RT-LAMP assay, what are the other subtypes of
influenza viruses (RNA samples) included in the study? (Enumerate)
H1N1, H3N2, H5N1, H5N6, H5N8, and H7N9

5. For Clinical evaluation, how many influenza-positive samples were used in the study?
73 influenza-positive

6. What is the sample collected from the patients? Where was is collected?
nasopharyngeal aspirate samples collected from patients at Chungbuk National University
Hospital, Korea

7. In clinical evaluation, the multiplex RT-LAMP was evaluated for specificity by testing it to
samples of other human infectious viruses other than those targeting influenza viruses. What
are these other viruses? (Enumerate)
1. HEV human enterovirus
2. , AdV adenovirus
3. , PIV parainfluenza virus
4. , MPV human metapneumovirus
5. , HboV human bocavirus, HRV human rhinovirus
6. , 229E human coronavirus 229E
7. , NL63 human coronavirus NL63
8. , OC43 human coronavirus OC43
9. , RSVA respiratory syncytial virus A
10. , RSVB respiratory syncytial virus B
11. , MERS Middle East respiratory syndrome coronavirus
8. In the results, what was the detection limit range of the RT-LAMP assay?
0.1 ~ 100 viral genome copies (p < 0.05)

9. How about the detection limit range for the conventional one-step RT-PCR and real-time RT-PCR
assay?
RT-PCR assays 0.1 ~ 1,000

10. Based on the result of the specificity of the RT-LAMP, was there cross-reactivity with other
viruses observed? Support your answer.
No cross-reactivity based on Table 3 because there are all negatives from all RT-LAMP samples

11. In the clinical samples, how many percents was the result for specifically detecting the following
influenza viruses by the multiplex RT-LAMP
Table 4
a. Seasonal influenza
b. Type B 100% (35/35)
c. H1N1 100%(12/12)
d. H3N2 97%(34/35)
e. H5N6 100%(8/8)
f. H5N8 100%(8/8)
g. H7N9 100%(1/1)

12. After reading the said paper or studying this topic, what are your
conclusions/thoughts/learnings/realization?

RT LAMP is a rapid test method than conventional RT-PCR

It also minimizes the use of expensive PCR Machine,also requires less labor
Thus, it is a simple, rapid and cheaper diagnostic method
RT-LAMP can be adapted to be used in other disease other than Influenza Virus