Supplementary webappendix This webappendix formed part of the original submission and has been peer reviewed. We post it as supplied by the authors. Supplement to: de Araújo TVB, Rodrigues LC, de Alencar Ximenes RA, on behalf of investigators from the Microcephaly Epidemic Research Group, the Brazilian Ministry of Health, the Pan American Health Organization, Instituto de Medicina Integral Professor Fernando Figueira, and the State Health Department of Pernambuco. Association between Zika virus infection and microcephaly in Brazil, January to May, 2016: preliminary report of a case-control study. Lancet Infect Dis 2016; published online Sept 15. http://dx.doi.org/10.1016/S1473-3099(16)30318-8.
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Supplementary webappendixThis webappendix formed part of the original submission and has been peer reviewed. We post it as supplied by the authors.
Supplement to: de Araújo TVB, Rodrigues LC, de Alencar Ximenes RA, on behalf of investigators from the Microcephaly Epidemic Research Group, the Brazilian Ministry of Health, the Pan American Health Organization, Instituto de Medicina Integral Professor Fernando Figueira, and the State Health Department of Pernambuco. Association between Zika virus infection and microcephaly in Brazil, January to May, 2016: preliminary report of a case-control study. Lancet Infect Dis 2016; published online Sept 15. http://dx.doi.org/10.1016/S1473-3099(16)30318-8.
Investigators contributing on behalf of: MERG Sergio Souza da Cunha ([email protected]), Carmen Dhalia ([email protected]), Marcela Santos ([email protected]), Fanny Cortes ([email protected]), Marcus K Eder ([email protected]) Brazilian Ministry of Health Wanderson Kleber de Oliveira ([email protected]), Giovanini Evelim Coelho ([email protected]) Pan American Health Organization Juan Jose Cortez-Escalante ([email protected]), Carlos Frederico Campelo de Albuquerque de Melo ([email protected]), Pilar Ramón-Pardo ([email protected]), Sylvain Aldighieri ([email protected]), Jairo Méndez-Rico ([email protected]), Marcos Espinal ([email protected]) IMIP Leuridan Torres ([email protected]), Adriano Nassri Hazin ([email protected]), Ana Van der Linden ([email protected]), Mônica Coentro ([email protected]) State Health Department George Santiago Dimech ([email protected]), Romildo Siqueira de Assunção ([email protected]), Patricia Ismael de Carvalho ([email protected]), Valdete Felix Oliveira ([email protected])
1
Annex Table 1: Line listing of cases and controls
n Micro-
cephaly Sex
Head circumference
(cm)
Brain Image
Maternal
Rash
Month of
gestation
Rash
Birth weight
(g)
Gestational
age
(week)
Small for gestational
age
Maternal
age
(years)
Maternal
schooling
(years)
Zika laboratory diagnosis
IgM
serum
(OD)
IgM
serum
result
RT-PCR
serum
Result
IgM
CSF
(OD)
IgM CSF
result
RT-PCR
CSF
result
RT-PCR or IgM
serum
RT-PCR or IgM
serum
or CSF
1 Case F 31·5 Normal Yes 7 2,955 39·4 Normal 16 8 1·0 Neg Neg - Not
collected
Not
collected Neg Neg
2 Case F 31·0 Normal Yes 9 2,435 39·0 SGA 23 11 0·6 Neg Neg 1·1 Neg Pos Neg Pos
3 Case F 28·0 Not available Yes 3 2,075 41·2 SGA 21 12 32·8 Pos Neg 15·0 Pos Pos Pos Pos
4 Case F 31·0 Normal No - 2,055 38·1 SGA 18 9 0·8 Neg Neg 24·2 Pos Neg Neg Pos
5 Case F 25·5 Abnormal Yes 3 2,325 40·2 SGA 17 10 13·4 Pos Neg 33·9 Pos Neg Pos Pos
6 Case M 28·0 Normal No - 1,660 34·5 SGA 32 3 1·1 Neg Neg 0·9 Neg Neg Neg Neg
7 Case F 28·0 Not available No - 2,565 39·4 SGA 17 10 13·6 Pos Neg 24·5 Pos Neg Pos Pos
8 Case F 29·6 Abnormal No - 1,520 36·1 SGA 42 9 0·9 Neg Neg 0·9 Neg Neg Neg Neg
9 Case M 29·0 Not available No - 1,955 37·2 SGA 24 12 - Not
collected
Not
collected -
Not
collected
Not
collected * Neg * Neg
10 Case M 31·5 Not available Don´t
know - 2,675 41·4 SGA 24 10 -
Not
collected
Not
collected -
Not
collected
Not
collected * Neg * Neg
11 Case F 27·4 Normal Yes 5 1,230 34·0 SGA 18 0 1·0 Neg Neg 1·1 Neg Neg Neg Neg
12 Case F 31·5 Normal Yes Don´t know
2,755 40.2 SGA 16 7 0·7 Neg Pos 0·6 Neg Pos Pos Pos
13 Case F 25·5 Normal Yes 1 1,080 33·0 SGA 30 10 0·7 Neg Neg 0·9 Neg Neg Neg Neg
14 Case F 32·0 Abnormal Yes 2 3,110 40·4 SGA 23 10 25·5 Pos Neg 37·1 Pos Neg Pos Pos
15 Case F 31·3 Abnormal No - 2,620 38·6 SGA 37 5 0·9 Neg Neg - Not
collected
Not
collected Neg Neg
16 Case F 27·0 Abnormal Yes 5 2,066 37·1 SGA 34 14 10·4 Pos Neg 23·8 Pos Neg Pos Pos
17 Case M 29·0 Abnormal No - 2,770 40·1 SGA 27 15 14·1 Pos Neg 20·5 Pos Neg Pos Pos
18 Case M 30·8 Abnormal Yes 3 2,930 40·3 Normal 16 10 16·6 Pos Neg 25·3 Pos Neg Pos Pos
19 Case F 30·5 Normal Yes 1 2,265 38·6 SGA 37 12 1·0 Neg Neg 0·9 Neg Neg Neg Neg
20 Case M 29·0 Normal No - 1,790 38·4 SGA 17 0 1·1 Neg Neg 1·1 Neg Pos Neg Pos
2
21 Case F 30·1 Normal No - 1,480 36·6 SGA 28 10 1·2 Neg Neg 1·1 Neg Neg Neg Neg
22 Case M 29·0 Normal No - 1,930 37·4 SGA 26 11 0·8 Neg Neg 0·8 Neg Neg Neg Neg
23 Case F 29·0 Normal No - 2,195 36·3 Normal 15 9 0·9 Neg Neg 1·2 Neg Neg Neg Neg
24 Case M 25·6 Not available No - 890 31·2 SGA 28 12 0·9 Neg Neg 0·9 Neg Neg Neg Neg
25 Case F 32·4 Normal Yes 8 2,395 40·5 SGA 24 12 0·9 Neg Neg 1·0 Neg Neg Neg Neg
26 Case F 31·5 Normal No - 3,310 39·2 Normal 28 7 1·0 Neg Neg - Not
collected
Not
collected Neg Neg
27 Case F 28·4 Normal No - 1,166 34·5 SGA 22 6 0·7 Neg Neg 0·9 Neg Neg Neg Neg
28 Case F 31·0 Abnormal No - 2,115 38·4 SGA 20 13 1·3 Neg Neg - Not
collected
Not
collected Neg Neg
29 Case F 31·0 Abnormal No - 2,315 39·3 SGA 24 12 1·2 Neg Neg 1·0 Neg Neg Neg Neg
30 Case M 30·0 Abnormal No - 2,950 39·5 Normal 27 13 17·1 Pos Neg 16·6 Pos Neg Pos Pos
31 Case M 31·5 Abnormal No - 2,125 40·4 SGA 18 12 0·6 Neg Neg 1·0 Neg Pos Neg Pos
32 Case F 31·7 Normal No - 2,350 39·4 SGA 15 9 0·7 Neg Neg - Not
collected
Not
collected Neg Neg
33 Control M 34·5 Normal No - 3,110 34·5 Normal 23 12 0·7 Neg Neg - - - Neg Neg
34 Control M 31·5 Normal No - 2,640 36·2 SGA 35 11 0·7 Neg Neg - - - Neg Neg
35 Control F 33·5 Normal No - 3,170 38·1 Normal 19 12 0·9 Neg Neg - - - Neg Neg
36 Control F 34·4 Normal Yes 8 2,652 38·2 Normal 22 0 0·7 Neg Neg - - - Neg Neg
37 Control F 35·0 Normal No - 2,995 38·5 Normal 22 10 1·0 Neg Neg - - - Neg Neg
38 Control F 33·0 Normal No - 2,730 36·3 Normal 33 10 1·2 Neg Neg - - - Neg Neg
39 Control F 33·0 Normal No - 2,680 36·4 Normal 16 12 1·1 Neg Neg - - - Neg Neg
40 Control M 34·0 Normal No - 3,180 39·0 Normal 42 12 0·9 Neg Neg - - - Neg Neg
41 Control F 34·5 Normal No - 2,860 37·6 Normal 33 12 0·9 Neg Neg - - - Neg Neg
42 Control M 35·0 Normal No - 3,525 40·6 Normal 18 10 0·9 Neg Neg - - - Neg Neg
43 Control M 35·0 Normal No - 3,400 39·4 Normal 24 12 1·1 Neg Neg - - - Neg Neg
44 Control F 42·5 Normal No - 3,415 38·5 Normal 27 8 0·8 Neg Neg - - - Neg Neg
45 Control F 35·0 Normal No - 3,545 38·3 Normal 35 10 0·9 Neg Neg - - - Neg Neg
46 Control F 33·0 Normal No - 3,375 40·0 Normal 17 9 0·9 Neg Neg - - - Neg Neg
3
47 Control F 35·0 Normal Don´t
know - 3,240 31·0 Normal 16 5 1·3 Neg Neg - - - Neg Neg
48 Control M 33·5 Normal Yes 9 3,045 39·1 Normal 19 5 - Not
collected Neg - - - Neg Neg
49 Control M 34·0 Normal No - 3,850 39·5 Normal 21 7 1·1 Neg Neg - - - Neg Neg
50 Control M 33·0 Normal No - 2,630 37·4 Normal 16 10 1·1 Neg Neg - - - Neg Neg
51 Control M 36·0 Normal No - 3,515 38·2 Normal 42 17 0·8 Neg Neg - - - Neg Neg
52 Control F 41·0 Normal No - 3,565 38·1 Normal 22 7 0·8 Neg Neg - - - Neg Neg
53 Control M 36·0 Normal No - 3,170 37·1 Normal 25 12 1·0 Neg Neg - - - Neg Neg
54 Control M 33·5 Normal Yes 8 3,590 38·0 Normal 19 11 1·3 Neg Neg - - - Neg Neg
55 Control M 35·0 Normal No - 2,850 39·4 Normal 28 12 1·0 Neg Neg - - - Neg Neg
56 Control F 34·5 Normal No - 3,560 38·6 Normal 21 9 0·7 Neg Neg - - - Neg Neg
57 Control M 35·0 Normal No - 3,685 39·3 Normal 41 12 0·9 Neg Neg - - - Neg Neg
58 Control F 36·0 Normal No - 3,530 38·3 Normal 16 6 1·0 Neg Neg - - - Neg Neg
59 Control F 33·0 Normal No - 2,745 39·3 Normal 24 9 0·7 Neg Neg - - - Neg Neg
60 Control F 33·5 Normal No - 2,385 35·6 SGA 21 12 0·7 Neg Neg - - - Neg Neg
61 Control M 35·0 Normal No - 3,295 39·3 Normal 19 11 0·8 Neg Neg - - - Neg Neg
62 Control F 34·5 Normal No - 2,720 37·1 Normal 16 11 1·0 Neg Neg - - - Neg Neg
63 Control F 33·5 Normal Yes 7 2,760 38·1 SGA 21 10 1·1 Neg Neg - - - Neg Neg
64 Control M 32·0 Normal No - 2,165 34·2 Normal 23 0 0·8 Neg Neg - - - Neg Neg
65 Control M 36·0 Normal No - 3,845 40·0 Normal 33 12 - Not
collected Neg - - - Neg Neg
66 Control F 36·0 Normal No - 3,530 38·2 Normal 25 11 1·1 Neg Neg - - - Neg Neg
67 Control M 36·0 Normal No - 4,020 40·5 Normal 25 11 0·8 Neg Neg - - - Neg Neg
68 Control M 32·5 Normal Yes 1 2,610 38·0 Normal 38 0 1·0 Neg Neg - - - Neg Neg
69 Control F 34·0 Normal No - 2,890 39·5 Normal 40 7 0·8 Neg Neg - - - Neg Neg
70 Control M 35·0 Normal Yes 7 3,295 37·3 Normal 20 7 1·0 Neg Neg - - - Neg Neg
71 Control F 31·8 Normal Yes 4 2,380 38·3 Normal 26 12 0·8 Neg Neg - - - Neg Neg
72 Control M 34·0 Normal No - 3,013 37·3 Normal 27 12 0·9 Neg Neg - - - Neg Neg
4
73 Control F 33·0 Normal Yes Don´t know
3,045 36·0 Normal 24 9 0·8 Neg Neg - - - Neg Neg
74 Control F 32·5 Normal No - 3,020 37·6 Normal 21 12 1·4 Neg Neg - - - Neg Neg
75 Control F 33·0 Normal Yes 6 2,970 41·4 Normal 34 5 2·1 Neg Neg - - - Neg Neg
76 Control F 35·0 Normal Yes 3 3,480 40·0 Normal 20 12 1·3 Neg Neg - - - Neg Neg
77 Control F 34·0 Normal No - 3,150 37·0 Normal 27 12 1·1 Neg Neg - - - Neg Neg
78 Control M 35·0 Normal No - 3,520 39·0 Normal 23 9 1·0 Neg Neg - - - Neg Neg
79 Control M 35·5 Normal No - 3,655 38·1 Normal 28 12 1·0 Neg Neg - - - Neg Neg
80 Control M 34·5 Normal No - 3,190 43·1 Normal 31 2 0·4 Neg Neg - - - Neg Neg
81 Control M 35·0 Normal Yes 8 3,615 39·0 Normal 18 9 1·6 Neg Neg - - - Neg Neg
82 Control M 33·0 Normal Yes 6 2,800 39·0 Normal 17 11 1·0 Neg Neg - - - Neg Neg
83 Control F 33·0 Normal Yes 6 2,840 40·0 Normal 33 10 - Not
collected Neg - - - Neg Neg
84 Control F 34·5 Normal No - 2,890 39·4 Normal 18 7 1·1 Neg Neg - - - Neg Neg
85 Control M 36·5 Normal No - 3,155 39·0 Normal 23 12 1·0 Neg Neg - - - Neg Neg
86 Control M 34·5 Normal No - 3,540 39·6 Normal 25 5 0·8 Neg Neg - - - Neg Neg
87 Control M 36·0 Normal No - 3,330 38·1 Normal 17 12 0·7 Neg Neg - - - Neg Neg
88 Control M 34·0 Normal No - 2,850 37·4 Normal 28 12 2·1 Neg Neg - - - Neg Neg
89 Control F 30·5 Normal Yes 7 2,360 35·6 SGA 32 13 0·8 Neg Neg - - - Neg Neg
90 Control F 37·0 Normal Yes 7 3,510 40·0 Normal 28 12 1·0 Neg Neg - - - Neg Neg
91 Control F 33·5 Normal No - 3,430 38·2 Normal 39 14 1·0 Neg Neg - - - Neg Neg
92 Control M 34·0 Normal No - 3,715 39·2 Normal 16 6 1·0 Neg Neg - - - Neg Neg
93 Control F 37·0 Normal No - 3,880 40·3 Normal 20 12 0·7 Neg Neg - - - Neg Neg
94 Control F 35·0 Normal No - 3,100 39·5 Normal 34 10 0·9 Neg Neg - - - Neg Neg
* Case 9 was stillbirth and case 10 was neonatal death, RT-PCR was tested in macerated tissues.
ELISA values are patient optical densities divided by negative control densities (OD); Values less than 2 were considered negative, 2–3 equivocal, and more than 3 positive;
CSF=cerebrospinal fluid.
5
Annex Table 2: Maternal ZIKV and DENV serotype specific antibodies (IgG) by plaque reduction
neutralization test (PRNT50)
Cases (n = 30)
N (%)
Controls (n = 61)
N (%)
Specificantibodies (PRNT50)*
ZIKV 5 (16·7) 4 (6·6)
ZIKV/DENV 19 (63·3) 35 (57·4)
ZIKV/DENV1/DENV2/DENV3/DENV4 0 1
ZIKV/DENV1/DENV3/DENV4 0 1
ZIKV/DENV3/DENV4 15 29
ZIKV/DENV3 3 3
ZIKV/DENV4 1 1
DENV 4 (13·3) 21 (34·4)
DENV3/DENV4 2 18
DENV1/DENV2 1 -
DENV3 1 3
Notdetected 2 (6·7) 1 (1·6)
* Not performed for 2 cases and 1 control
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