Supplementary Appendices:
Viral genome and antiviral drug sensitivity analysis of two
patients from a family cluster caused by the influenza A (H7N9)
virus in Zhejiang, China, 2013
HaiNv Gaoa,b, HangPing Yaoa,b, WeiFeng Lianga,b, XiaoXin Wua,b,
HaiBo Wua,b, NanPing Wua,b, ShiGui Yanga,b, Qiong Zhanga,b, KunKai
Sua,b, Jing Guoa,b, ShuFa Zhenga,b, YiXin Zhua,b, HongLin Chenb,c,
Kwok-yung Yuenb,c and LanJuan Lia,b.
aH.G., H.Y. ,W.L. and X.W. contributed equally to this work.
aState Key Laboratory for Diagnosis and Treatment of Infectious
Diseases, The First
Affiliated Hospital, College of Medicine, Zhejiang University,
No. 79 Qingchun Road, Hangzhou 310031, China;
bCollaborative Innovation Center for Diagnosis and Treatment of
Infectious Diseases, Hangzhou, China;
cState Key Laboratory for Emerging Infectious Diseases,
Department of Microbiology, The University of Hong Kong, Hong Kong
SAR, China
Correspondence: Lanjuan Li, MD, State Key Laboratory for
Diagnosis and Treatment of Infectious Diseases, First Affiliated
Hospital, College of Medicine, Zhejiang University, 79 Qingchun Rd,
Hangzhou, 310003, China ([email protected]).
Table of contents
Authors’ Contributions
P3
Normal ranges for all laboratory chemistries and definitions for
abnormal values
P4
Table S1 Type of exposure and total exposure time of the index
patient with the second patient
P5
Table S2 Epidemiology characteristic of a family cluster of two
laboratory-confirmed H7N9 virus infection patients
P6
Table S3 Clinical features and Laboratory Abnormalities of a
family cluster of two laboratory-confirmed H7N9 virus infection
patients
P7
Table S4 Complications, treatment and clinical outcomes of
patients infected with re-emerge avian-origin influenza A (H7N9)
virus
P9
Table S5 Nucleotide sequences identities among eight genes of
three isolates from two patients and one chicken epidemiological
linked to family cluster of two patients with influenza A (H7N9)
virus infection in Anji county, 2013
P10
Figure S1 Timeline of family cluster of two patients infected
with avian influenza A (H7N9) virus.
P12
Figure S2 The variation of chest radiological findings for two
epidemiological linked H7N9 virus infection patients
P13
Figure S3 Phylogenetic trees of the HA (A) (positions 19 to
1654) , NA (B) (positions 54 to 1371), PB2 (positions 43 to 2251),
PB1 (positions 90 to 2255), PA (positions 22 to 2063), NP
(positions 11 to 1485), M (positions 26 to 946), and NS (positions
188 to 817) genes of the H7N9 viruses isolated in a family cluster
cases.
P14
Authors’ Contributions
Drs. Lanjuan Li, KY Yuen designed the study. All the authors
have been taking care of the H7N9 cases and have been involved in
laboratory investigation. Drs. Hainv Gao, Hangping Yao Weifeng
Liang and Xiaoxing Wu analyzed the data. Dr. Hainv Gao wrote the
manuscript, Dr. Kunkai Su modified the manuscript. Drs. Hainv Gao,
Hangping Yao, Lanjuan Li and KY Yuen revised the manuscript. Drs.
Lanjuan Li decided to publish the paper.
Normal ranges for all laboratory chemistries and definitions for
abnormal values
Normal range of White cells was 4,000-10,000 per cubic
millimeter;
Normal range of Hemoglobulin for male was 12 -16 g/dl, for
female was 11-15 g/dl;
Normal range of C-reactive protein was less than 10
mg/liter;
Normal range of Procalcitonin was less than 0.5 ng/ml;
Normal range of Creatinine was less than 133 umol/liter;
Normal range of Lactate dehydrogenase was less than 250
U/liter;
Normal range of Creatine kinase was less than 200 U/liter;
Lymphocytopenia was defined as a lymphocyte count of less than
1500 per cubi millimeter;
Thrombocytopenia was defined as a platelet count of less than
150,000 per cubic millimeter;
Abnormal value of PaO2:FiO2 was defined as the ratio of less
than 300.
Table S1 Type of exposure and total exposure time of the index
patient with the second patient
Date
Exposure to the symptomatic patient
Type of exposure to the symptomatic patient
Total time of exposure
(h)
During the exposure time the patient had cough
Nov 19, 2013
-
Nov 20, 2013
＋
In the same room, talk with patient
0.3
＋
Nov 21, 2013
＋
In the same room, talk with patient
0.3
＋
Nov 22, 2013
＋
In the same room, talk with patient
0.5
＋
Nov 23, 2013
＋
In the same room, in the same car, talk with patient
2
＋
Nov 24, 2013
＋
In the same room, talk with patient
3
＋
Nov 25 2013
＋
In the same room, talk with patient, provide care to patient
10
＋
Nov 26, 2013
＋
In the same room, talk with patient, provide care to patient
18
＋
Nov 27, 2013
-
Table S2 Epidemiology characteristic of a family cluster of two
laboratory-confirmed H7N9 virus infection patients
The index patient
The second patient
Age (yr)
57
31
Date of illness onset
November 21, 2013
November 30, 2013
Sex
Male
Male
Occupation
Worker
Businessman
Date of laboratory confirmation of virus
November 25, 2013
December 5, 2013
Exposure to live poultry in previous 2 wks
＋
-
Exposure to symptomatic case patient in previous 2 wks
-
＋
Chronic Smoker
＋
-
Underlying disease
Any
＋
-
Chronic lung disease
-
-
Diabetes
-
-
Asthma
-
-
Hypertension
＋
-
Type of residence
Semiurban
Semiurban
BMI
26
25.4
Table S3 Clinical features and Laboratory Ab-rmalities of a
family cluster of two laboratory-confirmed H7N9 virus infection
patients
The index patient
The second patient
Presenting symptoms
Fever
＋
＋
Any cough
＋
＋
Productive cough
＋
＋
Hemoptysis
-
＋
Myalgia
＋
-
Shortness of breath
＋
＋
Gastrointestinal symptoms
-
＋
Diarrhea
-
＋
WBC (/mm3)
4100
3200
Lymphocyte (/mm3)
600
500
Lymphopenia
＋
＋
Platelet (/mm3)
160000
103000
Thrombocytopenia
-
-
CRP (mg/L)
160.1
34.1
PaO2/FiO2
52
300
Chest Radiology
Both Lungs Involvement
＋
-
Ground grass Opacity
＋
＋
Consolidation
＋
＋
Table S4 Complications, treatment and clinical outcomes of
patients infected with re-emerge avian-origin influenza A (H7N9)
virus
The index patient
The second patient
Characteristics
Complications
Pneumonia
＋
＋
ARDS
＋
-
Shock
＋
-
Acute Renal Failure
＋
-
Bacteria co-infection
-
-
Secondary infections
＋
-
Invasive Mechanical ventilation
＋
-
Antiviral drug
oseltamivir
paramivir
Time between initiation of antiviral therapy and the viral test
negative (days)
15
5
ECMO
＋
-
Artificial Liver Support System therapy
＋
-
Antibiotics
＋
＋
Any kind antibiotic
＋
＋
Combination therapy
＋
-
Corticosteroids
＋
-
IVIG
＋
-
Probiotics
＋
＋
Time between onset of symptoms and initiation of oseltamivir
(days)
4
5
Time between onset of symptoms and ARDS (days)
5
NA
Death
Yes
No
Table S5 Nucleotide sequences identities among eight genes of
three isolates from two patients and one chicken epidemiological
linked to family cluster of two patients with influenza A (H7N9)
virus infection in Anji county, 2013
Gene segment divergence
Percent identity
ZJU17
ZJU18
Chicken/ZJU06
PB2
ZJU17
−
99.9
96.0
ZJU18
0.1
−
96.1
Chincken/ZJU06
4.2
4.1
−
PB1
ZJU17
−
100.0
99.2
ZJU18
0.0
−
Chincken/ZJU06
0.9
0.9
−
PA
ZJU17
−
99.8
97.6
ZJU18
0.2
−
97.4
Chincken/ZJU06
2.5
2.7
−
NP
ZJU17
−
100.0
99.5
ZJU18
0.0
−
99.5
Chincken/ZJU06
0.5
0.5
−
HA
ZJU17
−
100.0
99.3
ZJU18
0.0
−
99.3
Chincken/ZJU06
0.7
0.7
−
NA
ZJU17
−
100.0
99.9
ZJU18
0.0
−
99.9
Chincken/ZJU06
0.1
0.1
−
M
ZJU17
−
100.0
97.4
ZJU18
0.0
−
97.4
Chincken/ZJU06
2.7
2.7
−
NS
ZJU17
−
100.0
94.4
ZJU18
0.0
−
94.4
Chincken/ZJU06
5.8
5.8
−
Figure S1 Timeline of family cluster of two patients infected
with avian influenza A (H7N9) virus.
The incubation period of the index case was 5 days, for the
secondary case was from 4 to 10 days. The secondary case has
history of closely contacting with the index case.
Figure S2 The variation of chest radiological findings for two
epidemiological linked H7N9 virus infection patients
The index patient
A1 (Day 3): CT shows unilateral Ground-Glass Opacities (GGO) and
slightly consolidation was observed on left lower lung on Day
3.
A2 (Day 5): CT shows rapid progress of GGO and consolidation of
both lungs.
The second patient
B1 (Day 3): CT shows GGO and partial consolidation was observed
on the lingual of left lung.
B2 (Day 5): CT shows GGO and progress consolidation in two
segments of left lung.
Figure S3 Phylogenetic trees of the HA (A) (positions 19 to
1654) , NA (B) (positions 54 to 1371), PB2 (positions 43 to 2251),
PB1 (positions 90 to 2255), PA (positions 22 to 2063), NP
(positions 11 to 1485), M (positions 26 to 946), and NS (positions
188 to 817) genes of the H7N9 viruses isolated in a family cluster
cases.
Sequences of H7N9 virus characterised in this study are
highlight by a circle. A/Zhejiang/DTID-ZJU17/2013 and
A/Zhejiang/DTID-ZJU18/2013 are human isolates and
A/chicken/Zhejiang/DTID-ZJU06/2013 is the epidemiologically linked
chicken isolate. The strains isolated in the first wave and in the
same county were listed in this tree, highlight by a circle too.
They are A/Zhejiang/DTID-ZJU01/2013, A/Zhejiang/DTID-ZJU10/2013 and
A/chicken/Zhejiang/DTID-ZJU01/2013 respectively. The scale bar
represents the distance unit between sequence pairs.
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