CO-INFECTION OF DENGUE VIRUS BY SEROTYPES 1 AND 4 IN PATIENT FROM MEDIUM SIZED CITY FROM BRAZIL

Colombo, Tatiana Elias; Vedovello, Danila; Mondini, Adriano; Reis, Andrea Francesli Negri; Cury, Amena Alcântara Ferras; Oliveira, Frank Hulder de; Cruz, Lilian Elisa Arão Antonio; Bronzoni, Roberta Vieira de Morais; Nogueira, Maurício Lacerda

doi:10.1590/S0036-46652013000400009

person Tatiana Elias Colombo

schoolFaculdade de Medicina de São José do Rio Preto (FAMERP), São José do Rio Preto, SP, BrazilschoolUniversidade Estadual Paulista “Júlio de Mesquita Filho” (IBILCE/UNESP), São José do Rio Preto, SP, Brazil

person Danila Vedovello

schoolFaculdade de Medicina de São José do Rio Preto (FAMERP), São José do Rio Preto, SP, Brazil

person Adriano Mondini

schoolUniversidade Estadual Paulista “Júlio de Mesquita Filho” (FCFAR/UNESP), Araraquara, SP, Brazil

person Andrea Francesli Negri Reis

schoolPrefeitura de São José do Rio Preto, Departamento de Vigilância em Saúde, São José do Rio Preto, SP, Brazil

person Amena Alcântara Ferras Cury

schoolPrefeitura de São José do Rio Preto, Departamento de Vigilância em Saúde, São José do Rio Preto, SP, Brazil

person Frank Hulder de Oliveira

schoolPrefeitura de São José do Rio Preto, Departamento de Vigilância em Saúde, São José do Rio Preto, SP, Brazil

person Lilian Elisa Arão Antonio Cruz

schoolPrefeitura de São José do Rio Preto, Departamento de Vigilância em Saúde, São José do Rio Preto, SP, Brazil

person Roberta Vieira de Morais Bronzoni

schoolFaculdade de Medicina de São José do Rio Preto (FAMERP), São José do Rio Preto, SP, Brazil

person Maurício Lacerda Nogueira

schoolFaculdade de Medicina de São José do Rio Preto (FAMERP), São José do Rio Preto, SP, Brazil

Correspondence to: Mauricio L. Nogueira, E-mail:
mnogueira@famerp.br

Financial Support: This work is supported by INCT-Dengue/CNPq and FAPESP (grant # 2012/11733-6 and 2011/10458-9 to MLN)

Figures (3)
Tables (1)

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Fig. 1 Electrophoresis in agarose gel showing the amplified products on two methodologies. PM = DNA ladder = 100 bp and 1 = Co-infection Sample RP/BR/507/2012, dengue 1 and 4 strains. (A) Lanciotti (1992) PCR assay showing the amplicon of 482 bp and 392 bp DENV-1 of DENV-4; (B) Bronzoni (2005) PCR assay showing the amplicon of 472 bp and 222 bp DENV-1 of DENV-4.

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Fig. 2 (A) Alignment of thirty-one amino acid sequences of Capsid Protein (C) of samples obtained at Genbank and the DENV-1 sequence isolated in this work. The strain RP/BR/2012/507a is shown in yellow and amino acid substitutions identified are marked in green. Only amino acids are shown as distinct from the reference sample (AB074760.1). (B) Alignment of thirty-three amino acid sequences of Capsid Protein (C) of samples obtained at Genbank and the DENV-4 sequence isolated in this work. The strain RP/BR/2012/507b is shown in yellow and amino acid substitutions identified are marked in green. The amino acid substitution F49L is marked in blue. Only amino acids are shown as distinct from the reference sample (JQ513335.1).

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Fig. 3 (A) Neighbor-Joining phylogenetic tree of 31 sequences of DENV-1 and the sequence RP/BR/2012/507a, inferred from Capsid gene (365 bp). RP/BR/2012/507a is highlighted in gray, the five DENV-1 genotypes are indicated with “}” and DENV serotypes 2, 3 and 4 are used as outgroups. “D” stands for “Dengue” and “1” represents the serotype. Genotype “G” is represented after the serotype, with the number of the genotype. Bootstrap values are shown for key nodes and the scale bar represents 0.05 nt changes per site. (B) - Neighbor-Joining phylogenetic tree of 33 sequences of DENV-4 and the sequence RP/BR/2012/507b, inferred from Capsid gene (257 bp). RP/BR/2012/507b is highlighted in gray, the three DENV-4 genotypes are indicated with “}” and DENV serotypes 1, 2 and 3 are used as outgroups. “D” stands for “Dengue” and “4” represents the serotype. Genotype “G” is represented after the serotype, with the number of the genotype. Bootstrap values are shown for key nodes and the scale bar represents 0.05 nt changes per site.

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Table 1 DENV-1 and DENV-4 sequences used for phylogenetic analysis, including serotype and genotype, isolation sites, isolation year, strain and GenBank accession numbers

Serotype and Genotype	Genbank Accession	Isolation site	Strains	Isolation Year
Serotype 1
D1G1	AB074760.1	Japan	Mochizuki	2001
D1G1	AF180817.1	USA	16007	1999
D1G1	JN054255.1	Sri Lanka	DV1_SL_2010b	2010
D1G1	GQ398255.1	Singapore	SG/07K3640DK1/2008	2008
D1G1	DQ193572.1	Fujian/China	Fj231/04	2004
D1G2	FJ196841.1	Guangzhou/China	GD54/03	2003
D1G2	FJ196842.1	Guangzhou/China	GD66/03	2003
D1G2	EF440432.1	Australia	ET243	2000
D1G2	EF025110.1	China	71/02GZ	2006
D1G3	AB074761.1	Japan	A88	2002
D1G3	DQ285561.1	Seychelles/ France	1480/04	2004
D1G4	U88535.1	Nauru Island	Clone WestPac	1997
D1G4	M23027.1	Nauru Island	Nauru Island	1994
D1G5	FJ850087.1	Northern Brazil	BR/BID-V2395/2006	2006
D1G5	AF226685.2	Rio de Janeiro/Brazil	Den1BR/90	2000
D1G5	AF513110.1	Curitiba/Brazil	BR/01-MR	2002
D1G5	FJ850075.1	Northern Brazil	BR/BID-V2381/2002	2002
D1G5	FJ850090.1	Northern Brazil	BR/BID-V2398/2007	2007
D1G5	FJ850084.1	Northern Brazil	BR/BID-V2392/2005	2005
D1G5	JX502769.1	Brazil	RR_DENV1_09007	2009
D1G5	FJ850070.1	Northern Brazil	BR/BID-V2374/2000	2000
D1G5	FJ850071.1	Northern Brazil	BR/BID-V2375/2000	2000
D1G5	AB519681.1	Brasilia/Brazil	SB 01057805 (DF02)	2002
D1G5	FJ850081.1	Northern Brazil	BR/BID-V2389/2004	2004
D1G5	HM043709.1	Brazil	55/2009ES	2009
D1G5	JN086990.1	Brazil	A	2010
D1G5	JN086991.1	Brazil	B	2010
Serotype 4
D4G1	FJ196850.1	Guangzhou/China	GD09/1990	1990
D4G1	AF289029.1	Guangzhou/China	Guangzhou B5	2000
D4G1	AF177542.1	Mindanao/Philippines	Mindanao/BDJ/1995	1995
D4G1	GQ868594.1	Philippines	PH/BID-V3361/1956/1956	1956
D4G2	EU854295.1	Puerto Rico	US/BID-V1083/1986	1986
D4G2	GQ868582.1	Santander/Colombia	CO/BID-V3409/2001	2001
D4G2	AH011968.1	Costa Rica	D4.108_1996CR	1996
D4G2	JN559741.2	Boa Vista/Brazil	ROR7365	2010
D4G2	JN983813.1	Boa Vista/Brazil	Br246RR/2010	2010
D4G2	HQ822126.1	Roraima/Brazil	Br347RR	2010
D4G2	HQ822125.1	Roraima/Brazil	Br272RR	2010
D4G2	JN092557.1	Brazil	SPH318527	2011
D4G2	JN712226.1	São José do Rio Preto/Brazil	RP/BR/2011/131	2011
D4G2	JN712225.1	São José do Rio Preto/Brazil	SJRP/BR/2011/167	2011
D4G2	JQ513335.1	Belém/Brazil	BeH778494	2011
D4G2	JN092556.1	Brazil	SPH319325	2011
D4G2	JN092555.1	Brazil	SPH319268	2011
D4G2	JN092556.1	Brazil	SPH319325	2011
D4G2	JQ513333.1	Boa Vista/ Brazil	ROR7542	2010
D4G2	JQ513340.1	Boa Vista/ Brazil	ROR7591	2010
D4G2	AY152252.1	San Juan/ Puerto Rico	D4.69_1987	1987
D4G2	AY152084.1	San Juan/ Puerto Rico	D4.84_1994	1994
D4G2	AY152056.1	San Juan/ Puerto Rico	D4.17_1998	1998
D4G2	AY152196.1	San Juan/ Puerto Rico	D4.28_1992	1992
D4G2	AY152336.1	San Juan/ Puerto Rico	D4M.5_1982	1982
D4G2	AY152360.1	Dominica	D4M.44_1981DM	1981
D4G3	HQ875339.1	Singapura	SG(EHI)D4/2641Y08	2008
D4G3	GQ398256.1	Singapura	SG/06K2270DK1/2005	2005
D4G3	AY618993.1	Bangkok /Thailand	ThD4_0734_00	2000
Serotypes 1 to 4 using as outgroups
DENV-1	AB074760.1	Japan		2001
DENV-3	AF317645.1	Guangxi/China		2000
DENV-2	HM181971.1	São Paulo/Brazil		2008
DENV-4	AF326573.1	Dominica		2000

CO-INFECTION OF DENGUE VIRUS BY SEROTYPES 1 AND 4 IN PATIENT FROM MEDIUM SIZED CITY FROM BRAZIL

Co-infecção por vírus dengue, sorotipos 1 e 4, em paciente de cidade de porte médio no Brasil

Abstracts