Although viruses are the most common cause of acute gastroenteritis (AGE) in humans, details about the causative viruses in AGE are largely unknown because many causative viruses are unable to be cultured by current culture techniques. In our study, fecal samples from 10 children under five years of age with unexplained AGE and 10 healthy children were investigated for RNA viruses using random priming (RP)-mediated sequence-independent single primer amplification (SISPA). The causative viruses in cases of cryptogenic diarrhea were then assessed for their potential diagnostic value. Of the 1,129 viral clones identified, rotavirus was most commonly associated with AGE (125 sequences, 22.4%). In contrast, bacteriophage was most common (43 sequences, 13.6%) in healthy children. The remaining 515 viral clones were unidentifiable. These findings suggest that investigation of cases or outbreaks of unexplained diarrhea using a metaviromic strategy is a new avenue for diagnosis.
Bacteriophages ; Child ; Clone Cells ; Culture Techniques ; Diagnosis ; Diarrhea ; Disease Outbreaks ; Feces* ; Gastroenteritis* ; Humans ; Parechovirus ; RNA Viruses ; Rotavirus

We report a human parechovirus-3 (HPeV-3) infection in 2 neonates who had prolonged fever (>5 days) with palmar-plantar erythema. This distinctive rash was observed 4–5 days after fever onset, just before defervescence. Elevated aspartate aminotransferase, lactate dehydrogenase, and ferritin levels were characteristic laboratory findings in the 2 cases, suggesting tissue damage caused by hypercytokinemia. Case 1 was treated with intravenous immunoglobulin, considering the possibility of severe systemic inflammatory responses. The initial ferritin level was 385 ng/mL (range, 0–400 ng/mL); however, the level increased to 2,581 ng/dL on day 5 after fever onset. Case 2 presented with milder clinical symptoms, and the patient recovered spontaneously. HPeV-3 was detected in cerebrospinal fluid and/or blood samples, but no other causative agents were detected. The findings from our cases, in accordance with recent studies, suggest that clinical features such as palmar-plantar erythema and/or hyperferritinemia might be indicators of HPeV-3 infection in neonates with sepsis-like illness. In clinical practice, where virology testing is not easily accessible, clinical features such as palmar-plantar erythema and/or hyperferritinemia might be helpful to diagnose HPeV-3 infection.
Aspartate Aminotransferases ; Cerebrospinal Fluid ; Erythema ; Exanthema ; Ferritins ; Fever ; Humans ; Immunoglobulins ; Infant, Newborn* ; L-Lactate Dehydrogenase ; Parechovirus ; Virology

OBJECTIVETo study HPeV from stool samples of children with acute gastroenteritis under 5 years old.

METHODSWe conducted a real-time PCR to detect HPeV from stool samples and to amply VP1 sequence by nested RT-PCR to identify HPeV type.

RESULTSThe results showed that 27 of 306 (8.82%) children with acute gastroenteritis were infected HPeV. 11 strains were typed. 9 strains HPeV1, both HPeV2 and HPeV4 was 1 strain. HPeV was mostly identified in autumn season with a peak in July. HPeV seemed relevant in children >2 years old. The range of nucleotide identity between all isolated strains with reference strains was 79%-92%.

CONCLUSIONEpidemiology characteristic of HPeV in Jilin was concordance with that of reports. HPeV3 wasnt detected. It's significant to conduct the large scale and long-term surveillance of HPeV.

Acute Disease ; Child, Preschool ; Female ; Gastroenteritis ; epidemiology ; virology ; Humans ; Infant ; Male ; Parechovirus ; classification ; genetics ; isolation & purification ; Phylogeny

Human parechovirus type 3 (HPeV3) is an important pathogen of severe sepsis. HPeV3 is a non- enveloped, single-stranded, positive-sense RNA virus with a linear and continuous genomic RNA. The complete genome of a HPeV3 (BJ-C3174) strain was analyzed from the serum specimen from a child with sepsis hospitalized in Beijing, China, in 2012. The whole genome of BJ-C3174 was 7329 nucleotides (nt) in length excluding a poly (A) tail. One large open reading frame (ORF) of 6531 nt encoding a putative polyprotein precursor of 2177 amino acids (aa) was flanked by a 5' untranslated region (UTR) of 709 nt and 3' UTR of 91 nt. Phylogenetic analysis showed that BJ-C3174 belonged to HPeV3 and was closest to the HPeV3 strain BONN-2 from Germany. Compared with HPeV1-8 reference strains, BJ-C3174 shared the highest similarities with BONN-2 in full length and in each of the gene segments of the genome. The nucleotide and predicted amino acid identities of the whole genome between BJ-C3174 and BONN-2 were 99.3% and 99.8%, respectively, which were higher than those compared with HPeV3 prototype. Recom- bination of the gene segment with other HPeVs types was not identified.
Amino Acid Sequence ; Child ; Genome, Viral ; Humans ; Molecular Sequence Data ; Parechovirus ; classification ; genetics ; Phylogeny ; Sepsis ; blood ; virology

PURPOSE: This study was conducted to evaluate epidemiological data of the viral pathogens obtained from stool exams and provide information on the regional prevalence of infectious diarrheal disease west in Gyeonggi Province, Korea. METHODS: We enrolled a cohort of children <10 years of age admitted for treatment of acute diarrhea at Bucheon St. Mary's Hospital, College of Medicine, The Catholic University of Korea. In total, 310 fecal specimens, documented to be free of common bacterial pathogens, were collected from pediatric patients during a 12-month period from January to December 2009 and were tested for the presence of rotavirus, parechovirus, adenovirus, astrovirus, enterovirus, and norovirus using polymerase chain reaction (PCR) and reverse transcription polymerase chain reaction (RT-PCR) assay. RESULTS: The most common virus was parechovirus (16%), followed by adenovirus (15%), astrovirus (14%), rotavirus (13%), and enterovirus (5%). Interestingly, only one of the specimens was positive for norovirus. Single infection cases were detected in 173 (55.8%) of the 310 children, whereas mixed viral infections were detected in 10 (3.2%) of the same children. Viral gastroenteritis generally showed a double peak of incidence. Parechovirus, rotavirus, and adenovirus shared a similar pattern of peak incidence with overall viruses; however, astrovirus infections occurred more frequently in the spring. Eighty-five percent of the confirmed viral gastroenteritis cases developed in under 24 months. CONCLUSION: The results support the importance of parechovirus, adenovirus, astrovirus, and enterovirus as causative agents of diarrhea in children, which may be underestimated by current routine diagnostic testing.
Adenoviridae ; Child ; Child, Hospitalized ; Cohort Studies ; Diagnostic Tests, Routine ; Diarrhea ; Dysentery ; Enterovirus ; Gastroenteritis ; Humans ; Incidence ; Korea ; Norovirus ; Parechovirus ; Polymerase Chain Reaction ; Prevalence ; Reverse Transcription ; Rotavirus ; Viruses

OBJECTIVETo detect human parechovirus (HPeV) from stool samples of hospitalized children for acute gastroenteritis of undetectable etiology.

METHODSWe conducted a real-time PCR to detect HPeV.

RESULTThe results showed that 24 of 99 (24%) children with gastroenteritis of undetectable etiology were detected with HPeV. Four known HPeV types (HPeV1, 3, 4, 6) were detected in the present study. HPeV1 (50%) was frequently identified as the predominant strain and follow by HPeV3 (25%), HPeV4 (8.3%) and HPeV6 (4.2%). We were unable to type 3 samples.

CONCLUSIONHPeV was prevalent in hospitalized children for acute gastroenteritis of undetectable etiology in China. Further study is needed for clarifying the role of HPeV in gastroenteritis.

Child, Preschool ; Feces ; virology ; Female ; Gastroenteritis ; virology ; Humans ; Infant ; Infant, Newborn ; Male ; Molecular Sequence Data ; Parechovirus ; classification ; genetics ; isolation & purification ; Phylogeny ; Picornaviridae Infections ; virology

PURPOSE: Human parechovirus (HPeV) is an increasingly recognized pathogenic cause of central nervous system (CNS) infection in neonates. However, HPeV infections have not been studied in older children. This study determined the prevalence and clinical features of HPeV CNS infection in children in Korea. METHODS: Reverse transcription polymerase chain reaction assays were performed using HPeV-specific, 5′ untranslated, region-targeted primers to detect HPeV in cerebrospinal fluid (CSF) samples from children presenting with fever or neurologic symptoms from January 1, 2013, to July 31, 2014. HPeV genotyping was performed by sequencing the viral protein 3/1 region. Clinical and laboratory data were retrospectively abstracted from medical records and compared with those of enterovirus (EV)-positive patients from the same period. RESULTS: Of 102 CSF samples, six (5.9%) were positive for HPeV; two of 21 EV-positive samples were co-infected with HPeV. All samples were genotype HPeV3. Two HPeV-positive patients were <3 months of age and four others were over 1 year old. While HPeV-positive infants under 1 year of age presented with sepsis-like illness without definite neurologic abnormalities, HPeV-positive children over 1 year of age presented with fever and neurologic symptoms such as seizures, loss of consciousness, and gait disturbance. The CSF findings of HPeV-positive patients were mostly within the normal range, whereas most (73.7%) EV-positive patients had pleocytosis. CONCLUSIONS: Although HPeV is typically associated with disease in young infants, the results of this study suggest that HPeV is an emerging pathogen of CNS infection with neurologic symptoms in older childhood.
Central Nervous System Infections* ; Central Nervous System* ; Cerebrospinal Fluid ; Child ; Enterovirus ; Fever ; Gait ; Genotype ; Humans* ; Infant ; Infant, Newborn ; Korea ; Leukocytosis ; Medical Records ; Neurologic Manifestations ; Parechovirus* ; Polymerase Chain Reaction ; Prevalence ; Reference Values ; Retrospective Studies ; Reverse Transcription ; Seizures ; Unconsciousness

PURPOSE: Human parechovirus (HPeV) and enterovirus (EV) are causative agents of a sepsis-like illness in neonates and of infections of the central nervous system in young children. The objectives of this study were to assess the prevalence of HPeV3 and EV infection in young children with a sepsis-like illness or with meningitis in Jinju, Korea. METHODS: Cerebrospinal fluid (CSF) samples were collected from 267 patients (age range, 1 day to 5 years) and assessed for HPeV and EV by performing reverse transcription polymerase chain reaction assay. Amplification products of the VP3/VP1 region of HPeV and of the VP1 region of EV were sequenced to identify the virus type. RESULTS: HPeV and EV were detected in 3.4% and 7.5% of the total CSF samples assessed, respectively. The age distribution of EV-positive patients (median age, 1.4 months) had a significantly broader range than that of HPeV-positive patients (median age, 7.8 months). The peak seasons for HPeV and EV infection were spring and summer, respectively. The clinical symptoms for HPeV and EV infection were similar, and fever was the most common symptom. Pleocytosis was detected in 22.2% of HPeV-positive patients and 35.5% of EV-positive patients. The VP3/VP1 gene sequence of the nine Korean strains clustered most closely with the Japanese strain (AB759202). CONCLUSION: The data indicate that HPeV infection is predominant in young infants (<6 months) and that meningitis without pleocytosis was caused by both HPeV and EV infection in children.
Age Distribution ; Asian Continental Ancestry Group ; Central Nervous System ; Cerebrospinal Fluid* ; Child* ; Enterovirus* ; Fever ; Gyeongsangnam-do ; Humans ; Infant ; Infant, Newborn ; Korea ; Leukocytosis ; Meningitis ; Parechovirus* ; Polymerase Chain Reaction ; Prevalence* ; Reverse Transcription ; Seasons

Child ; Herpesviridae ; classification ; Herpesviridae Infections ; diagnosis ; Humans ; Paramyxoviridae ; classification ; Paramyxoviridae Infections ; diagnosis ; Parechovirus ; classification ; Pediatrics ; Picornaviridae ; classification ; Picornaviridae Infections ; diagnosis ; SARS Virus ; classification ; Severe Acute Respiratory Syndrome ; diagnosis ; Virus Diseases ; diagnosis ; Viruses ; classification

OBJECTIVEHuman parechovirus (HPeV) is a single-stranded, positive sense RNA virus in the Parechovirus genus within the large family of Picornaviridae. As a possible new pathogen of neonatal sepsis, meningoencephalitis and other infections in young children, HPeV gets more and more attention. This study aimed to better understand the association of HPeV with central nervous system (CNS) infectious diseases and sepsis among hospitalized children in Beijing.

METHODA total of 577 cerebrospinal fluid (CSF) samples were retrospectively collected from 557 children suspected of CNS infections in 2012. Three hundred and fifty-one of them were male and 206 were female. HPeV was screened by reverse transcription-nested PCR (RT-nPCR) with the universal primers which target the highly conserved 5'UTR. The positive samples were genotyped by amplifying and sequencing for the VP3/VP1 junction region. The sequences were compared with the HPeV sequences from GenBank and performed phylogenetic analysis.Some samples other than CSF from HPeV positive children, including serum, nasopharyngeal aspirate and stool, were collected and carried out screening for HPeV.

RESULTWith the RT-nPCR by universal primers, HPeVs were detected in 18 out of 577 CSF samples obtained from 18 children with a positive rate of 3.1%. The ratio of male and female was 2: 1. There were no statistically significant differences on infection rate between boys (12/351, 3.4%) and girls (6/206, 2.9%). All of 18 positive CSF samples were negative for enterovirus, Epstein-Barr virus (EBV), human cytomegalovirus (HCMV), and herpes simplex virus 1 and 2 (HSV).HPeVs from 10 positive CSF samples were genotyped successfully, consisting of 7 HPeV3 and 3 HPeV1. In addition, 2 of 8 serum samples were positive for HPeV3 and 1 of 2 stool samples were positive for HPeV 1. HPeVs were identified in CSF from children aged from 15 days to 14 years, in which 7 cases were infants younger than 3 months and 5 cases were infants from 3 months to one year. Three children older than the age of 9 years (9, 13 and 14 years) were positive for HPeV. Most of the children (6/8) infected with HPeV3 were younger than 3 months and were diagnosed as sepsis, while the rest of HPeV3 positive children were diagnosed as meningitis and bronchopneumonia. HPeV3 infection clustered in August, while HPeV1 in January.

CONCLUSIONHPeVs were associated with CNS infections and sepsis in hospitalized children in Beijing, especially in children younger than one year.HPeV3 was the predominant type identified in CSF.

Adolescent ; Age Distribution ; Central Nervous System Infections ; cerebrospinal fluid ; epidemiology ; virology ; Cerebrospinal Fluid ; virology ; Child ; Child, Hospitalized ; Child, Preschool ; Female ; Genotype ; Humans ; Infant ; Infant, Newborn ; Male ; Parechovirus ; classification ; genetics ; isolation & purification ; Picornaviridae Infections ; cerebrospinal fluid ; epidemiology ; virology ; RNA, Viral ; genetics ; Retrospective Studies ; Reverse Transcriptase Polymerase Chain Reaction ; Seasons ; Sepsis ; cerebrospinal fluid ; epidemiology ; virology ; Sequence Analysis, DNA