Identiﬁcation and genetic characterization of two porcine
astroviruses from domestic piglets in China
Received: 12 February 2015 / Accepted: 8 August 2015 / Published online: 4 September 2015
Ó Springer-Verlag Wien 2015
Abstract Porcine astrovirus are divided into ﬁve geno-
types. In this study, we identiﬁed two porcine astroviruses
(AstV-LL-1 and AstV-LL-2) by using sequence-indepen-
dent single-primer ampliﬁcation (SISPA) on faecal speci-
mens of healthy domestic piglets younger than 15 days.
The detection rate for both was 2.82 % (14/497). AstV-LLs
were then sequenced and characterised. Phylogenetic
analysis revealed that they have the characteristics of
porcine astrovirus (PastV) 2 and 5 and have some unique
genetic features. Our ﬁndings show that the two astro-
viruses are novel lineages of PAstV2 and 5. The ﬁndings
may be helpful in evaluating the ecology and evolution of
astroviruses in pigs.
Keywords Porcine astrovirus Á Domestic pig Á Faeces
Astroviruses are small, non-enveloped viruses with posi-
tive-sense single-stranded RNA that belong to the family
Astroviridae, which is divided into two genera, Mamas-
trovirus and Avastrovirus, which infect mammals and
birds, respectively . Currently, ﬁve distinct genotypes of
porcine astrovirus (PAstV) have been characterized. All
ﬁve porcine astroviruses have been detected in faecal
samples from healthy pigs [8, 9, 13]. Additionally, PAstV4
has been identiﬁed in both domestic pigs and in wild boars
. PAstV2 and PAstV4 have also been detected in blood
from healthy pigs of different ages . PAstV3 was
detected in a faecal sample from a pig suffering from
diarrhoea . Several outbreaks of porcine-associated
zoonotic diseases have occurred in recent years, including
outbreaks of Nipah virus  and Streptococcus suis .
With the rapid development of stock breeding in China, the
potential threat of zoonotic diseases spreading to humans is
growing; therefore, further research is needed to evaluate
astrovirus ecology and its evolution in pigs.
This report describes the identiﬁcation and genetic
characterization of two novel lineages of PAstV2 and
PAstV5 from healthy domestic piglets in China.
In 2006, 497 porcine stool samples from young healthy
piglets (\15 days of age) were collected from several
farms in Lulong County, China. All samples were stored at
-80 °C until use.
The protocol used to identify novel PAstVs with an
E-value cutoff of 10
was adapted from our previous
study . Two pooled faecal samples from 10 different
pigs were ampliﬁed to produce two sequence-independent
single-primer ampliﬁcation (SISPA) products, which yiel-
ded 85 clones from one and 150 clones from the other. All
235 clones were sequenced and aligned against the NCBI
Nr database using the BLASTn and BLASTx algorithms.
The 85 clones from the ﬁrst sample matched with viruses,
bacteria, phages, eukaryotic cells, and unknown sources
with constituent ratios of 7 %, 46 %, 11 %, 29 %, and
7 %, respectively. Of the six viral sequences
(E \ 1 9 10
in BLAST analysis), ﬁve matched other
viruses with a high similarity (2 enteroviruses, 2 rota-
viruses, and 1 sapovirus), and one sequence, which we
named AstV-LL-1, showed 82 % identity to PAstV
J. Li, M. Li and L. Zheng contributed equally to the manuscript.
Electronic supplementary material The online version of this
article (doi:10.1007/s00705-015-2569-x) contains supplementary
material, which is available to authorized users.
& Zhao-jun Duan
National Institute for Viral Disease Control and Prevention,
China CDC, 100 Ying-Xin St., Xuan-Wu District,
Beijing 100052, China
Arch Virol (2015) 160:3079–3084