How epidemiology and
diagnostics drive public policy-lessons from influenza
Public Health Practice Grand Rounds
Johns Hopkins Bloomberg School of Public Health
Charlotte A. Gaydos, MS, MPH, Dr PH
Professor
Division of Infectious Diseases
Johns Hopkins University
Baltimore, Maryland, USA
Objectives
1. Requirements for rapid point-of-care
(POC) tests for influenza in real time
2. Limitations of some Rapid POC tests
3. Overview of some new POC tests
4. Overview of a new rapid PLEX-ID
technology which can identify and
genotype influenza samples for just- intime surveillance needs
Requirements for rapid point-of-care
(POC) tests for influenza in real time
1. Sensitive
2. Specific
3. Fast
4. Simple
5. Inexpensive
ASSURED: WHO
recommendations:
a.
b.
c.
d.
e.
f.
g.
Affordable
Sensitive
Specific
User Friendly
Rapid and Robust
Equipment-Free
Deliverable to End-Users
Current Types of Tests for Influenza and
Respiratory Viruses
Rapid Tests
insensitive, non-specific
DFA
specific
rapid, less sensitive,
Shell Vial culture
sensitive
Roller Tube culture
sensitive, slow
Real time qPCR/RT-PCR
sensitive, slow
Pyrosequencing
expensive
Flow through microsphere array
rapid, expensive
Positivity Rates for 3 Rapid
Influenza Diagnostic Tests
Rapid Flu Dx Test
Influenza A Type
Binax NOW Influenza
A&B Novel H1N1
Seasonal H1N1
H3N2
40%
60 %
80%
(18/45)
(3/5)
(12/15)
Directigen EZ Flu A+B
Novel H1N1
Seasonal H1N1
H3N2
49%
75%
83%
(21/43)
(3/4)
(10/12)
QuickVue A+B
Novel H1N1
Seasonal H1N1
H3N2
69%
80%
80%
(31/45)
(4/5)
(12/15)
CDC, MMWR August 17, 2009
% Positive (#+/# tested)
Performance of the QuickVue
Rapid Test for Influenza
NHRC Influenza Testing: 4/20/01 – 5/30/09
3066 Total Specimens Processed
Confirmed S-OIV
273 (8.9%)
Seasonal H3N2
31 (2.0%)
Seasonal H1N1
18 (0.61%)
QuickVue Rapid Test (Quidel) results available for 767 patients
#Pos by QuickVue
#Pos by PCR
Sensitivity
95% CI
S-OIV
20/39
52%
35% - 67%
Seasonal H1N1
12/19
63%
39% - 82%
Seasonal H2N3
6/19
31%
14% - 57%
Influenza Type
Specificity of the test, compared to RT-PCR, was 99% in all cases
Faix D et al. N Engl J Med 2009;10.1056/NEJMc0904264
Rapid Antigen Test for Diagnosis of
Pandemic (Swine) Influenza A/H1N1 (N=84)
Rapid Influenza Diagnostic Test
Sensitivity Specificity
Binax NOW Influenza A&B
38.3%
100%
BD Directigen EZ Flu A+B
46.7%
100%
QuickVue Influenza A+B
53.3%
100%
Vasoo et al. CID 2009;49:1090-1093
Evaluation of Multiple test methods for
Detection of Novel 2009 H1N1 in NYC
(N=1831)
Influenza Diagnostic Test
Rapid Ag Test
Sensitivity Specificity
21.2%
99.5%
DFA
47.2%
99.6%
R-Mix
98.49%
100%
(BinexNOW or 3M Rapid Detection)
Ginocchio et al. J Clin Vir 2009;45:191-195
Influenza:
Always a challenge; always changing
H1N1pdm 2009
H3N2v 2011-12
New influenza H3N2v
•Influenza A (H3N2) variant (H3N2v) virus
infection was were detected in the United States
during July 2011–July 2012
H3N2 influenza A with H1N1pdm09 M (matrix) gene
• In 2012, 306 cases of H3N2v infections were
reported to CDC from 13 states (12 in 2011 in 5)
IN (138 cases), OH (106), WS (20), MD (12), PA (11),
MI (6), IL & MN (4), WV (3), HI & UT (1)
~93% occurred in persons aged <18 years, 16
hospitalized, 1 death
•All reported direct or indirect exposure to swine at fairs
New variant: Minnesota Health: reports that the number
of H1N2v cases has increased to 4
New influenza H3N2v
• H3N2v viruses can be detected by qualified U.S. public
health laboratories using the CDC Flu rRT-PCR Dx Panel.
Initially, if specimens tested positive for influenza A, H3,
and pandemic influenza A markers and negative for H1 and
pandemic H1 markers, they were reported as inconclusive.
•On August 7, CDC updated the results interpretation of
the CDC Flu rRT-PCR Dx Panel for H3N2v for public health
laboratories. Specimens with these findings may now be
reported as “presumptive positive for influenza A (H3N2)v
virus” and, for the ongoing investigations, cases with
presumptive-positive test results at the state or local
public health laboratory will now be classified as
confirmed, as are those cases confirmed at CDC.
•The CDC Flu rRT-PCR Dx Panel is available in public
health laboratories but is not a point-of-care test available
to clinicians
Evaluation of seven FDA-cleared RIDTs for the
ability to detect H3N2v viral antigens — CDC, 2012
RIDT
(manufacturer)
Name
Approved
Specimens
Analyzer
interpret
BinaxNOW Influenza
A&B (Alere)
BinaxNOW
NP swab
Nasal wash/
aspirate/swab
No
Directigen EZ Flu A+B
(Becton-Dickinson
Directigen
NP wash/Aspirate /swab No
throat swab
SAS FluAlert A&B
(SA Scientific)
FluAlert
Nasal wash/aspirate
No
QuickVue Influenza A+B
Test (Quidel)
QuickVue
NP swab Nasal wash
/aspirate/swab
No
Sofia Influenza A+B
(Quidel)
Sofia
NP aspirate/swab/wash
Nasal wash
Required
BD Veritor System for
Veritor
Rapid Detection of Flu
A+B (Becton) Dickinson)
NP swab/nasal swab
Required
Xpect Flu A&B
(Remel)
Nasal wash/swab
Throat swab
No
Xpect
Number of 10-fold virus dilutions (maximum =
five) detected by seven FDA-cleared RIDTs, by
H3N2v strain designation — CDC, 2012
Strain
TCID
50/ml
Binax Direct Flu
NOW -ogen Alert
Quick Sofia Veritor
View
Xpect
A/Kansas
/12/2009
104.5
1
4
U
U
2
4
4
A/Pennslylv
14/2012
104.5
2
4
U
2
2
4
3
A/Minnesota
/11/2012
104.5
U
3
U
U
3
3
2
A/Indiana
/08/2011
106.0
1
3
U
U
2
3
2
A/Indiana
/10//2011
104.0
U
3
U
U
2
4
2
A/West VA
/06/2011
106.0
2
3
U
2
4
4
2
A/Iowa
/07/2011
104.5
2
4
1
1
3
4
3
CDC Recommendations
•The fact that a negative RIDT result should not be
considered as conclusive evidence of lack of infection
with influenza A (H3N2)v
•Results from RIDTs, both positive and negative, always
should be interpreted in the broader context of the
circulating influenza strains present in the area, level of
clinical suspicion, severity of illness, and risk for
complications in a patient with suspected infection
• Clinicians should minimize the occurrence of false RIDT
results by strictly following the manufacturer’s
instructions, collecting specimens within the first 72 hours
of illness, and confirming RIDT results by sending a
specimen to a public health laboratory, who will send to
CDC
Additional CDC guidance available at http://www.cdc.gov/flu/swineflu/h3n2vtesting.htm.
Two new amplified flu POC tests
GeneXpert® Flu, Cepheid
FilmArray, Idaho Technology
GeneXpert® Flu
No Requirement for dedicated PCR area
CLIA Moderate
Complexity
Flocked swab or Nasal
Asp/wash
3. Binding reagent into port 1
• Results in ~ 1 hr
• POC, random access
3
Xpert® Flu Assay
Result Interpretation
• Xpert Flu provides
display of:
– Flu A Positive
– Flu A Positive with
2009 H1N1 Detected
or
– Flu B Positive
• Reporting of 2009
H1N1 requires
presence of both
Flu A and 2009
H1N1 targets
Xpert® Flu Assay
Published Data
Denver Children’s Hospital:
 Xpert® Flu is a highly
accurate.
 No prior molecular training
required.
 Clinically actionable
Prodesse ProFlu+
Sensitivity
Xpert Flu
(Flu A)
Xpert Flu
100%
100%
100%
97.9%
99.5%
(2009 H1N1)
University of Virginia:
Prodesse ProFlu+
 Xpert® Flu is a rapid, robust,
sensitive, highly specific
and user-friendly test
99.1%
(Flu B)
Xpert Flu
Sensitivity
Xpert Flu
Xpert Flu
for 24hr test availability
Xpert Flu
(2009 H1N1)
Specificity
97.3%
100%
94.4%
100%
100%
100%
(Flu A)
(Flu B)
 Reduced complexity allows
Specificity
Gen Xpert Fu amplified assay
compared to rapid influenza tests
Influenza A Influenza A Influenza
B
Sensitivity Specificity Sensitivity
97.3%
100%
100%
Xpert Flu
Direct
Fluorescent 95.9%
Antibody
Influenza
B
Specificity
100%
99.2%
100%
100%
Binax NOW 62.2%
100%
54.5%
100%
BD
Directogen
100%
48.5%
100%
71.6%
*Sensitivity and specificity were calculated using rRT-PCR as the comparator
DiMaio et al. J of Vir Meth, 2012
Film Array
Comparison of positive and negative results by
FilmArray RP and Prodesse assays for viruses
detectable by both assays
Virus
No. of specimens
Sens % Spec %
Adenovirus
Influenza A
(H1-2009)
Influenza B
Metapneumo
Parainflu 1
Parainflu 3
RSV
PRO-/FA+ PRO+/FA+ P RO+/FA- PRO-/FA+
181
6
5
0
191
191
180
187
179
181
1
1
12
3
10
10
0
0
0
0
0
0
0
0
0
2
3
1
54.5
100
100
100
100
100
100
100
100
100
100
98.9
98.4
99.5
Prodesse assays were considered the gold standard for comparison.
Viruses targeted by both FilmArray RP and Prodesse assays, but not detected during this
study: influenza A H1 seasonal virus, influenza A H3 seasonal virus, and parainfluenza virus
2
PRO, Prodesse; FA, FilmArray Respiratory Panel
Loeffelholtz et al. JCM 49:4083-4088, 2012
Sensitivity and specificity of the FilmArray
RP (FA) and xTAG RVP (xTAG) versus PCRconfirmed results
v
Virus
No. detecte
Film Assay Sens % Spec %
FA+, xTAG+ FA+, xTAG- FA-, xTAG+ FA-, xTAG-
Influ A
32
0
1
Influ B
7
0
0
RSV
37
8
0
155
100
100
Parainflu
15
1
0
184
100
100
Rhino/Entero 39
4
2
155
95.6 100
Adenovirus
9
0
190
90
Metapneumo
6
193
100
1
Rand et al. JCM 49:2449-2453, 2011
1
0
167
193
97
100
100
100
99.5
100
Comparison of the FilmArray System
to Real-Time PCR for Detection of
Respiratory Pathogens in Children
•215 frozen archived pediatric respiratory specimens
previously characterized as either negative or positive
for one or more pathogens by real-time PCR were
examined using the FilmArray RP
•Agreement between FilmArray RP and real-time PCR
assays for shared analytes was 98.6% (kappa0.92 )
The combined positive percent agreement was 89.4%;
the negative percent agreement was 99.6%
Positive % agreement
•Influenza A virus H1
Influenza A virus H3
Influenza A virus 2009 H1
Influenza B virus
100
88.2
94.1
88.2
Negative % Agreement
100
100
100
100
Pierce et al. JCM 50:364-371, 2012
Recent Reviews
Chartrand et al. Annals Inter Med 156:500-511,
2012
Kumar at al. Clin Microbiol Rev 25:344-361,
2012
Vijayan et al. Emerg Infect Dis 18:1414-1421,
2012
Evaluation of Sofia
fluorescent immunoassay for
Influenza A/B virus
Sofia FIA displayed sensitivities of 82.2% and
77.8% for strains A and B respectively,
compared to the RT-PCR
Sensitivities of BinaxNOW A/B antigen kit, and
Directigen Flu A and B were 54.8%, and 68.5%
for influenza A
Sensitivities of BinaxNOW A/B antigen kit, and
Directigen Flu A and B were 62.5%, and 52.8%
for influenza B
Lee et al. J Clin Vir 2012 (in press)
How Do We Do Surveillance?
Challenges for Influenza Typing and
Surveillance for Respiratory Viruses
•Epidemics happen quickly: 4wk H1N1pdm
•Called Type A influenza: “not H3” and “not
H1”, thus “un-subtypeable”
•*Challenge to have a surveillance tool that
gives a rapid diagnosis, as well as a
genotype, that can provide public health
guidance for “just-in-time” diagnostics
*Metzgar, D. et al. J Clin Microbiol. 48:4228-4234., 2010
PLEX-ID as a New Surveillance Tool
• PLEX-ID Technology combines broad
amplification with PCR, and electrospray
ionization mass spectrometry
• Designed to provide broad identification,
detailed genotyping and
characterization, and recognition of
known and emerging strains
• Designed to track a potential pandemic
in real time
•Comprehensive sample tracking
•Analysis of 300+ samples in 24 hours
•Simplified, easy to interpret results
Respiratory Virus Surveillance I Assay
• Influenza virus validation at Ibis
• >650 blinded samples
–Nasal aspirates, nasal swabs, nasal washes,
throat swabs, bronchial washes, tracheal
aspirates
–Samples from 1999-2006, across United
States
–Correctly identified all Influenza A types
• 149 H3N2
• 34 H1N1
Influenza
–67 Influenza B
Sensitivity
96.8%
Sampath. R., K.L. Russell, C. Massire, M.W. Eshoo, V.
Harpin, L.B. Blyn, et al. Rapid Surveillance of the Global
Spread of Emerging Influenza Virus Genotypes by Mass
Spectrometry. PLoS ONE. May 2007, issue 5, e489.
Specificity
PPV
NPV
97.5%
96.0%
98.0%
Novel Influenza Virus: 2009 H1N1
Before the H1N1 pandemic strain was added to the PLEX
database, it appeared as a hybrid of several strains from
different hosts – human, avian, and swine
Human
Avian
Swine
Pandemic Influenza Samples from AprilJune 2009; Deyde et al. Plos One Oct 2010 5:e13293
CDC ID
T5000 Inferred Subtype
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
PB1
NP
M1
PA
NS1
NS2
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
39 32 24 33
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
24 28 24 29
36 25 27 24
36 25 27 24
35 26 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
35 26 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 25 27 24
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
36 35 20 28
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
34 29 16 26
Comparison to
CDC ID
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Pandemic Influenza Samples Johns Hopkins
University October-December 2009
CDC ID
T5000 Inferred
Subtype
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
Swine-H1N1
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
A/California/04/2009
PB1
NP
M1
PA
NS1
NS2
38 33 25 32
38 33 25 32
38 33 25 32
39 32 24 33
38 33 25 32
38 33 25 32
38 33 25 32
38 33 25 32
37 34 25 32
37 34 25 32
37 34 25 32
37 34 25 32
38 33 25 32
38 33 25 32
38 33 25 32
39 32 24 33
38 33 25 32
39 32 24 33
39 33 24 32
37 34 25 32
38 33 25 32
38 33 25 32
38 33 25 32
38 33 25 32
37 34 25 32
39 32 24 33
38 33 25 32
38 33 25 32
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
34 22 21 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
35 21 20 25
24 28 24 29
23 29 25 28
24 28 24 29
23 29 25 28
22 30 25 28
23 29 25 28
23 29 25 28
24 28 24 29
23 29 25 28
24 28 24 29
24 28 24 29
23 29 25 28
23 29 25 28
23 29 25 28
23 29 25 28
23 29 25 28
23 29 25 28
24 28 24 29
23 29 25 28
24 28 24 29
23 29 25 28
24 28 24 29
24 28 24 29
24 28 24 29
23 29 25 28
23 29 25 28
24 28 24 29
24 28 24 29
22 30 25 28
24 28 24 29
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
38 25 29 20
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 26 28 23
35 35 21 28
35 35 21 28
35 36 21 27
35 36 21 27
35 35 21 28
35 36 21 27
35 36 21 27
35 35 21 28
35 36 21 27
35 36 21 27
35 36 21 27
35 36 21 27
35 35 21 28
35 36 21 27
35 35 21 28
35 35 21 28
35 36 21 27
35 35 21 28
35 35 21 28
35 35 21 28
35 36 21 27
35 36 21 27
35 36 21 27
35 35 21 28
35 36 21 27
35 36 21 27
35 36 21 27
35 36 21 27
35 36 21 27
35 35 21 28
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
34 29 16 26
33 30 17 25
34 29 16 26
33 30 17 25
33 30 17 25
33 30 17 25
34 29 16 26
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
33 30 17 25
Comparison to
CDC ID
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Matched
Conclusions
• Know your Rapid POC test, its strengths and
limitations
• Some POC tests may not work well with new
strains of influenza
• Promising new POC tests are becoming available
• The PLEX-ID Influenza Surveillance Typing Assay
can detect newly assorted and shifted or drifted
strains (including avian strains such as H5 and
other H and N types)
• Be aware of the potential of new influenza strains
Questions?
Acknowledgements
Kuan-Fu Chen, MD
[email protected]
855 N Wolfe St
530 Rangos Bldg.
Baltimore, MD 21205
PH 410-614-0932
FAX 410-614-9775
Results of RT-PCR Analyses of 3066
Specimens, According to Influenza A Subtype
Faix D et al. N Engl J Med 2009;10.1056/NEJMc0904264
FilmArray RP Pouch Pathogens, Gene Targets
and LOD95
Organism
Gene Target(s)
Strain
LOD95
vAV
Hexon
Type 1
BoV
NP-1
Clinical Sample
B. per
Toxin
A639
C. pne
ompA
TW183
CoV 229E
Polymerase
VR-740
CoV HKU1
Nucleoprotein
PCMC 6123
CoV OC43
Nucleoprotein
VR-759
CoV NL63
Nucleoprotein
NR-470
EV
5’ UTR
Echovirus 6
hMPV
Nucleoprotein
hMPV-16/IA10-2003 Type A1
HRV
5’UTR
1A
Flu A (H1N1) Matrix, NS1, HA1
A/Brisbane/59/07
Flu A (H1N1- 2009) Matrix, NS1, HA1-2009 A/SwineNY/03/2009
Flu A (H3N2) Matrix, NS1, HA3
A/Wisconsin/67/2005
Flu B
Hemagglutinin
B/FL/04/06
M. pne
Toxin
M129 – Type 1
PIV 1
Hemagglutinin
Type 1
PIV 2
Fusion
Type 2
PIV 3
Fusion
Type 3
PIV 4
Fusion
Type 4a
RSV
Matrix
RSV Type A
Poritz et al. P:os One e26047, 2011
300
4000
4,000
3000
4
1.96 x 10^6
600
5
30,000
2
1
200
100
5
60
30
500
10
10
5,000
2
What circulated in 2010-2111 in Baltimore?
Pandemic 2009 H1N1 strains
#
Influenza A/New York/15/2009
16
Influenza A/California/05/2009 pandemic-H1N1
7
Influenza A/Santo Domingo/WR1068N/2009(Pandemic-H1N1)PB1-S4 SNP
3
Influenza A virus A/Singapore/GP2711/2009(Pandemic-H1N1)NUC-S2 SNP
1
Influenza A/Wisconsin/629-D00698/2009(Pandemic-H1N1)-HAS1 SNP
1
Influenza A/Hiroshima/220/2009(Pandemic-H1N1)
1
Influenza A/New York/4057/2009(Pandemic-H1N1)-NS1-S1 SNP
2
Influenza A/Japan/PR1070/2009(Pandemic-H1N1)-PA-S3 SNP
1
Murillo et al. ICEID 2012
What circulated in 2010-2111 in Baltimore?
H3N2 strains
#
Influenza A/Thailand/CU-B1697/2009
9
Influenza A/ Kentucky/UR07-0148/2008
8
Influenza A/Boston/47/2008(H3N2)
1
Influenza A/New York/3079/2009(H3N2)
1
Influenza A/Texas/AF01/2007(H3N2)
1
Influenza
A/California/VRDL334/2009(H3N2)
1
Influenza A/Cheong/2009
1
Murillo et al. ICEID 2012