Detection and Genetic Characterization of Isolates of Hepatitis E Virus from Pigs and Human in Chungnam Region of Korea

Mi-Kyung Chae; Ji-Young Kim; Jiaqi Chu; Duck-Hwan Kim; Han Sang Yoo; Chang-Sik Park; Myung-Cheol Kim; Moo-Hyung Jun

doi:10.4167/jbv.2006.36.1.31

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Chae, Kim, Chu, Kim, Yoo, Park, Kim, and Jun: Detection and Genetic Characterization of Isolates of Hepatitis E Virus from Pigs and Human in Chungnam Region of Korea

Original Article

J Bacteriol Virol 2006;36(1):31-39.

Published online: 18 February 2006

DOI: https://doi.org/10.4167/jbv.2006.36.1.31

Detection and Genetic Characterization of Isolates of Hepatitis E Virus from Pigs and Human in Chungnam Region of Korea

Mi-Kyung Chae, Ji-Young Kim, Jiaqi Chu, Duck-Hwan Kim, Han Sang Yoo¹, Chang-Sik Park², Myung-Cheol Kim, Moo-Hyung Jun^∗,

College of Veterinary Medicine, Chungnam National University, Daejeon 305-764, Korea

¹College of Veterinary Medicine, Seoul National University, Seoul 138-200, Korea

²Research Center for Transgenic Cloned Pigs, Chungnam National University, Daejeon 305-764, Korea

^∗Corresponding author: Moo-Hyung Jun. College of Veterinary Medicine, Chungnam National University, 220 Gungdong, Youseongu, Daejeon 305-764, Republic of Korea. Phone: +82-42-821-6753, Fax: +82-42-821-6753 e-mail: mhjun@cnu.ac.kr

Received 2 February 2006 Accepted 22 March 2006

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Swine hepatitis E virus (HEV) has been reported as a new zoonotic agent due to its close genomic resemblance to the human HEV. Recently this virus is indicated as one of the important pathogens in xenotransplantation that uses pig as a donor animal. We carried out to investigate the prevalence of HEV infections among the pigs and human population in Chungnam region using a nested RT-PCR for detection of a part of HEV ORF2 gene. The sequences of the amplified DNA were analyzed and the genetical divergency were characterized. A total of 18 HEV strains, comprising 16 strains from pig and 2 strains from human, were genetically isolated from the fecal and serum samples. Among the isolates, 5 strains (2.5%) were detected from 200 swine sera and 2 strains (2.0%) from 100 human sera. All of the 16 swine strains were isolated from the pigs at 3 month of age, but none of age groups revealed the positive for swine HEV RNA. In comparison of the nucleotide sequence between 16 swine HEV and 2 human HEV isolates, the range of identities was 91.5% to 100%. Two human HEV isolates shared 99.7% homology. In phylogenetic analysis, all of the isolates were classified into genotype III, and the 18 isolates were also closely related to the prototype of swine HEV and human HEV strains isolated in the United States and others recently identified from swine in Japan and Netherland.

Keywords: Hepatitis E virus, Swine HEV, Human HEV, Nucleotide sequences

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Figure 1.

Phylogenetic tree analysis of Korean isolates of swine HEV by comparison of the conserved 323 bp (6020∼6342 nt) of ORF2 in the various swine and human HEV isolates. Groups; I, II, III and IV.

Table 1.

Detection of HEV RNA in the specimens from the pigs and humans

Species	Age groups	No. of samples tested	No. of positive samples (%)		Designation of isolates
Species	Age groups	No. of samples tested	Feces	Serum	Designation of isolates
Pig	2^a 3 4 5 >7 (sows) Subtotal	29 67 49 25 30 200	0 11 (16.4) 0 0 0 11 (5.5)	0 5 (7.5) 0 0 0 5 (2.5)	SWKB1F, SWKB2F, SWKB3F, SWKB4F, SWKB5F, SWKB6F, SWKB7F, SWKB1S, SWKB2S, SWKB3S, SWKB4S, SWKB5S SWKT1F, SWKT2F, SWKT3F, SWKY1F
Human	30∼50^b	100	NT	2 (2.0)	KorHEV-1, KorHEV-2

^a Month old,

^b Years old, NT; not tested

Table 2.

Pairwise comparison of the nucleotide identity of partial ORF2 gene (6020∼6342 nt) of 18 HEV isolates identified in this study and other selected strains of HEV worldwide^a

Isolate		% identity with isolate no.
No	Name	1	2	3	4	5	6	7	8	9	10	11	12	13	14	15	16	17	18	19	20	21	22	23	24	25	26	27	28	29	30	31	32	33	34	35	36	37	38	39	40
1	Egypt		89.8	76.8	79.9	78.3	89.8	78.6	88.2	78.6	77.7	78.0	77.7	75.2	87.6	77.7	88.5	76.2	87.6	78.9	78.6	51.4	76.5	77.4	78.0	77.1	77.1	78.0	77.1	77.7	76.8	78.0	76.5	77.7	78.3	77.1	76.8	77.7	78.0	76.5	76.8
2	Hev037			77.1	78.9	78.3	89.2	77.7	91.6	77.7	78.6	78.9	79.3	78.0	89.8	79.3	92.0	77.1	91.3	78.0	77.7	51.4	76.2	78.6	78.3	78.3	78.3	78.9	78.0	78.6	77.7	78.9	77.4	78.6	78.6	78.3	77.7	78.6	78.9	77.4	77.7
3	HEV-US1				79.3	89.2	78.0	87.0	77.1	87.0	93.8	93.5	92.0	77.7	77.1	91.6	77.4	90.7	76.8	80.2	80.2	52.3	73.4	92.6	92.6	91.6	92.3	93.5	92.3	93.2	92.0	93.5	91.6	93.2	93.2	92.3	92.0	93.2	93.5	91.6	92.0
4	Ch-T21					79.9	79.6	79.3	79.9	79.3	78.9	79.3	78.9	85.4	80.8	78.3	80.2	77.4	79.9	87.3	87.9	251.1	77.7	79.3	78.9	78.9	78.9	79.3	78.6	78.9	78.3	79.3	78.6	78.9	79.3	78.9	78.3	78.9	79.3	78.6	78.3
5	JRA1						78.3	91.0	77.4	91.0	89.8	89.5	87.9	77.4	80.5	87.3	78.9	87.0	76.5	79.6	80.5	52.0	74.3	86.7	87.0	87.3	86.7	89.5	87.6	89.2	87.3	89.5	87.6	89.2	89.2	86.7	87.3	89.2	89.5	87.6	87.3
6	Moroco							77.4	89.8	77.4	78.0	78.3	78.6	75.9	86.7	78.0	87.6	78.0	89.2	80.2	79.3	52.6	76.5	79.6	79.9	79.6	79.3	78.3	77.4	78.0	77.1	78.3	77.4	78.0	78.6	79.3	77.1	78.0	78.3	77.4	77.1
7	NLSW20								78.0	100.0	87.9	87.6	86.1	77.4	78.3	85.4	77.4	84.8	77.1	79.3	79.6	54.5	74.9	85.1	85.1	85.8	84.8	87.6	85.8	87.3	85.4	87.6	85.1	87.3	87.9	84.8	85.4	87.3	87.6	85.1	85.4
8	Sar-55									78.0	78.0	78.3	77.7	78.0	90.1	78.0	92.9	77.7	97.5	78.0	80.2	52.3	77.7	77.7	77.4	77.7	77.4	78.3	77.4	78.0	77.1	78.3	76.8	78.0	78.0	77.4	77.1	78.0	78.3	76.8	77.1
9	swj681										87.9	87.6	86.1	77.4	78.3	85.4	77.4	84.8	77.1	79.3	79.6	54.5	74.9	85.1	85.1	85.8	84.8	87.6	85.8	87.3	85.4	87.6	85.1	87.3	87.9	84.8	85.4	87.3	87.6	85.1	85.4
10	swKOR-1											99.7	94.4	78.0	77.1	93.5	77.4	91.6	77.7	79.6	79.6	50.8	73.7	93.2	93.2	96.6	92.9	99.7	97.8	99.4	97.5	99.7	97.2	99.4	99.4	92.9	97.5	99.4	99.7	97.2	97.4
11	swKOR-2												94.7	77.7	76.8	93.8	77.1	92.0	78.0	79.9	79.9	51.1	74.0	93.5	93.5	96.9	93.2	100.0	98.1	99.7	97.8	100.0	97.5	99.7	99.7	93.2	97.8	99.7	100.0	97.5	97.8
12	swKOR-3													78.6	78.0	91.6	77.7	90.4	76.8	79.3	78.6	51.7	74.6	93.2	93.2	93.5	92.9	94.7	92.9	94.4	92.6	94.7	92.3	94.4	94.4	92.9	92.6	94.4	94.7	92.3	92.6
13	Tw32sw														77.4	76.8	77.1	76.8	76.8	83.3	83.9	52.3	73.1	76.8	77.1	77.7	76.5	77.7	76.5	77.4	76.2	77.7	76.5	77.4	77.7	76.5	76.2	77.4	77.7	76.5	76.2
14	Vietnam															78.0	96.6	76.5	90.1	77.1	78.0	50.8	77.7	77.1	76.8	76.8	76.8	76.8	75.2	76.5	74.9	76.8	75.2	76.5	76.5	76.8	74.9	76.5	76.8	75.2	74.9
15	HEV-US2																77.1	93.2	78.3	79.3	78.6	49.5	72.4	92.3	92.3	92.6	92.0	93.8	92.0	93.5	91.6	93.8	91.3	93.5	94.1	92.0	91.6	93.5	93.8	91.3	97.6
16	Burma																	75.9	93.5	77.7	79.3	51.1	78.3	76.8	76.5	77.1	76.5	77.1	76.2	76.8	75.9	77.1	75.5	76.8	76.8	76.5	75.9	76.8	77.1	75.5	75.9
17	USswineHEV																		78.0	78.3	77.7	51.4	74.9	92.0	92.3	92.0	91.6	92.0	90.1	91.6	89.8	92.0	89.5	91.6	91.6	91.6	89.8	91.6	92.0	89.5	89.8
18	Hetian																			77.7	79.9	52.0	78.3	78.0	77.7	78.0	77.7	78.0	77.1	77.7	76.8	78.0	76.5	77.7	77.7	77.7	76.8	77.7	78.0	76.5	76.8
19	JSN-Sap																				96.9	52.3	76.8	77.4	77.7	78.9	77.1	79.9	78.6	79.6	78.3	79.9	78.0	79.6	79.9	77.1	78.3	79.6	79.9	78.0	78.3
20	swj7–1																					53.6	77.4	76.8	77.1	78.3	76.5	79.9	78.6	79.6	78.3	79.9	78.0	79.6	79.9	76.5	78.3	79.6	79.9	78.0	78.3
21	avian																						49.8	5.08	51.1	51.7	50.8	51.1	50.2	51.4	50.2	51.1	49.8	50.8	51.1	50.8	50.2	50.8	51.1	49.8	50.2
22	Mexico																							74.6	74.3	74.9	74.9	74.0	72.8	73.7	72.4	74.0	72.1	73.7	73.7	74.9	72.4	73.7	74.0	72.1	72.4
23	SWKB-1F																								98.8	96.0	99.1	93.5	91.6	93.2	91.3	93.5	91.0	93.2	93.2	99.1	91.3	93.2	93.5	91.0	91.3
24	SWKB-1S																									96.0	98.5	93.5	91.6	93.2	91.3	93.5	91.0	93.2	93.2	98.5	91.3	93.2	93.5	91.0	91.3
25	SWKB-2F																										96.3	96.9	95.0	96.6	94.7	96.9	94.4	96.6	96.6	96.3	94.7	96.6	96.9	94.4	94.7
26	SWKB-2S																											93.2	91.3	92.9	91.0	93.2	90.7	92.9	92.9	100.0	91.0	92.9	93.2	90.7	91.0
27	SWKB-3F																												98.1	99.7	97.8	100.0	97.5	99.7	99.7	93.2	97.8	99.7	100.0	97.5	97.8
28	SWKB-3S																													98.1	99.7	98.1	99.4	97.8	97.8	91.3	99.7	97.8	98.1	99.4	99.7
29	SWKB-4F																														97.8	99.7	97.5	99.4	99.4	92.9	97.8	99.4	99.7	97.5	97.8
30	SWKB-4S																															97.8	99.7	97.5	97.5	91.0	100.0	97.5	97.8	99.7	100.0
31	SWKB-5F																																97.5	99.7	99.7	93.2	97.8	99.7	100.0	97.5	97.8
32	SWKB-5S																																	97.2	97.2	90.7	99.7	97.2	97.5	100.0	99.7
33	SWKB-6F																																		99.4	92.9	97.5	99.4	99.7	97.2	97.5
34	SWKB-7F																																			92.9	97.5	99.4	99.7	97.2	97.5
35	SWKT-1F																																				91.0	92.9	93.2	90.7	91.0
36	SWKT-2F																																					97.5	97.8	99.7	100.0
37	SWKT-3F																																						99.7	97.2	97.5
38	SWKY-1F																																							97.5	97.8
39	KorHEV-1																																								99.7
40	KorHEV-2

^a References for other HEV strains used in comparisons: Hev037 (X98292, India), Sar-55 (M80581, Pakistan), Vietnam (AF170450), Burma (M73218), JRA1 (AP003430, Japan), Hetian (L08816, China), Ch-T21 (AF151963, China), Tw32sw (AF117280, Taiwan), Morocco (AF065061), Mexico (M74506), U.S. swine HEV (AF082843), HEV-US1 (AF060668, U.S.), HEV-US2 (AF060669, U.S.), avian (AY043166, U.S.). swj7–1 (AB094219, Japan), JSN-Sap (AB091395, Japan), swj681 (AB073910, Japan), NLSW20 (AF336290, Netherlands), Egypt (AF051351), swKOR1 (AF516178, Korea), swKOR2 (AF516179, Korea), and swKOR3 (AF527942, Korea)

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