Human Bocavirus Genotypes Research Articles

Molecular characterization of human bocavirus in recycled water and sewage sludge in Thailand.

The study aimed to assess the presence and molecular characterization of human bocavirus (HBoV) in recycled water and sewage sludge samples in Thailand. One hundred and two recycled water and eighty-six sewage sludge samples collected from a wastewater treatment plant were tested for the presence of HBoV using nested PCR with broad-range primer pairs targeting the capsid proteins VP1 and VP2. HBoV DNA was detected in recycled water of 9/102 (8.8%) samples and sewage sludge of 27/86 (31.4%) samples. Based on DNA sequencing and phylogenetic analysis, the HBoV DNA sequences had 98.8-100.0% nucleotide identity to the sequences from HBoV reported globally. Thirty-five HBoV-positive samples were identified to genotypes as the predominant HBoV2; 26 followed by HBoV3; 8 and the rare HBoV4; 1 sample. Concerning recycled water, HBoV2 was detected in 3 (2.9%) and HBoV3 was detected in 5 (4.9%) of all samples. The sewage sludge samples were characterized as HBoV2 in 23 (26.7%), HBoV3 in 3 (3.5%) and HBoV4 in 1 (1.2%) of all samples. The frequency of HBoV detected in recycled water and sewage sludge samples significantly differed in sample type (p-value=0.007). The findings of three HBoV genotypes in recycled water and sewage sludge emphasized the circulation of the virus in the environment and the potential source of transmission to the community.

Prevalence and Molecular Characterization of Human Bocavirus Detected in Croatian Children with Respiratory Infection.

Human bocavirus (HBoV) 1 is considered an important respiratory pathogen, while the role of HBoV2-4 in clinical disease remains somewhat controversial. Since, they are characterized by a rapid evolution, worldwide surveillance of HBoVs’ genetics is necessary. This study explored the prevalence of HBoV genotypes in pediatric patients with respiratory tract infection in Croatia and studied their phylogeny. Using multiplex PCR for 15 respiratory viruses, we investigated 957 respiratory samples of children up to 18 years of age with respiratory tract infection obtained from May 2017 to March 2021 at two different hospitals in Croatia. Amplification of HBoV near-complete genome or three overlapping fragments was performed, sequenced, and their phylogenetic inferences constructed. HBoV was detected in 7.6% children with a median age of 1.36 years. Co-infection was observed in 82.2% samples. Sequencing was successfully performed on 29 HBoV positive samples, and all belonged to HBoV1. Croatian HBoV1 sequences are closely related to strains isolated worldwide, and no phylogenetic grouping based on mono- or co-infection cases or year of isolation was observed. Calculated rates of evolution for HBoV1 were 10−4 and 10−5 substitutions per site and year. Recombination was not detected among sequences from this study.

Molecular Quantification of Human Bocavirus in Environmental Water Samples in Giza, Egypt

There are extremely limited data on the prevalence and quantification of human bocavirus (HBoV) genotypes in different environments, particularly for rivers and drinking water. Here we performed quantitative real-time PCR (qPCR) to investigate the presence and load of HBoV genotypes in Nile water, drinking water, and effluent of the activated sludge wastewater treatment facility in Giza, Egypt. The results revealed that the detection rate of HBoV was 95.8% (23/24) and 29.2% (14/48) in effluents of the wastewater treatment facility and Nile water samples, respectively. HBoV was not found in drinking water samples from the Giza governorate. The mean concentration values of human bocavirus genotype HBoV-1, HBoV-2/4 and HBoV-3 were 0.52 log10 GC/L, 2.2 log10 GC/L, 0.3 log10 GC/L, respectively in HBoV-positive Nile water samples (n = 14), while in HBoV-positive effluent of wastewater samples (n = 23) were 0.77 log10 GC/L, 2.3 log10 GC/L, and 1.6 log10 GC/L, respectively. The viral type had a significant impact on the prevalence of the virus in surface water as well as effluent samples. A seasonal pattern of HBoV subtypes was observed in surface water (P = 0.0002), as the highest detection rate was found during winter. Furthermore, the discharge of the effluent with higher viral concentration into the water sources leads to viral contamination of these sources, which may be a probable source for human infections. This study is executed to fill a gap of knowledge concerning the quantification and genotyping of bocavirus in Nile water, and enrich the scientific literature body by providing data about HBoV in environmental samples.

Human bocavirus genotypes 1, 2, and 3 circulating in pediatric patients with acute gastroenteritis in Chiang Mai, Thailand, 2012–2018

BackgroundGlobal distribution of human bocavirus (HBoV) has been known to associate with viral gastroenteritis in pediatric population. This study was conducted in Chiang Mai, Thailand from 2012 to 2018 to investigate epidemiology and genotype distribution of HBoV in pediatric patients less than 5 years old hospitalized with diarrhea. MethodsA total of 2727 fecal specimens were investigated for the presence of HBoV using nested-PCR targeting partial VP1 capsid region. The detected HBoV strains were further characterized by nucleotide sequencing and phylogenetic analysis. ResultsDetection rate of HBoV infection in pediatric patients with acute diarrhea was 5.2%. Three genotypes of HBoV were detected with the most predominance of HBoV1 (50.4%), followed by HBoV2 (42.5%), and HBoV3 (7.1%). The majority of HBoV positive cases were children of 1 to <2 years old (31.9%) with high detection rate of HBoV1 and HBoV2. HBoV infection occurred all year-round. Phylogenetic analysis revealed that majority of HBoV1 displayed the genetic relationship with HBoV1 strains reported previously from Asia whereas only a few were related to the strains from Europe, South America, and Middle East. The HBoV2 and HBoV3 were also mainly closely related to the strains reported from Asia and a few from South America and North Africa. ConclusionsThis study highlights distribution of HBoV genotypes circulating in pediatric patients with acute gastroenteritis in Chiang Mai, Thailand. Overall, three genotypes of HBoV were detected with equally high prevalence of HBoV1 and HBoV2 whereas HBoV3 was detected with much lower prevalence.

Human Bocavirus in Brazil: Molecular Epidemiology, Viral Load and Co-Infections.

Human bocavirus (HBoV) is an emerging virus and has been detected worldwide, especially in pediatric patients with respiratory and gastrointestinal infection. In this study, we describe HBoV prevalence, genotypes circulation and DNA shedding, in stool samples from children up to two years of age in Brazil. During 2016 and 2017, 886 acute gastroenteritis (AGE) stool samples from ten Brazilian states were analyzed by TaqMan®-based qPCR, to detect and quantify HBoV. Positive samples were genotyped by sequencing the VP1/2 overlap region, followed by phylogenetic analysis and co-infections were accessed by screening other gastroenteric viruses. HBoV was detected in 12.4% (n = 110) of samples, with viral load ranging from 1.6 × 102 to 1.2 × 109 genome copies per gram of stool. From these, co-infections were found in 79.1%, and a statistically lower HBoV viral load was found compared to viral loads of rotavirus, norovirus and adenovirus in double infected patients (p < 0.05). No significant differences were found between HBoV viral load in single or co-infections, age groups or genotypes. Phylogenetic analysis identified the circulation of HBoV-1 in 38%, HBoV-2 in 40% and HBoV-3 in 22%. Continuous HBoV monitoring is needed to clarify its role in diarrhea disease, especially in the absence of classic gastroenteric viruses.

Human Bocavirus genotypes 1 and 2 detected in younger Amazonian children with acute gastroenteritis or respiratory infections, respectively

ObjectivesThis study aimed to verify the frequency, genotypes, and etiological role of Human Bocavirus (HBoV) in younger Amazonian children with either acute gastroenteritis (AGE) or respiratory infections (ARI). The influence of Rotarix™ vaccination and co-infection status was also investigated. DesignHBoV quantitative polymerase chain reaction (qPCR) testing was done on both fecal and saliva (1468 samples) from 734 children < 5 months old living in the Amazon (Brazil, Guyana, and Venezuela). High and median HBoV viral load samples were used for extraction, nested PCR amplification, and sequencing for genotyping. HBoV mRNA detection was done by reverse transcription following DNA amplification. ResultsThe overall HBoV frequencies were 14.2% (69/485; AGE) and 14.1% (35/249; ARI) (p = 0.83). HBoV exclusively infected 4.5% (22/485; AGE) and 4% (10/249) of the Amazonian children (Odds ratios 1.13, 95% confidence interval= 2.42-0.52). HBoV 1 was mainly detected in feces and saliva from AGE children; and HBoV2, from ARI children. HBoV mRNA was detected only in feces. The Rotarix™ vaccination status did not affect the HBoV frequencies. ConclusionsWe suggest that, after entry into the air/oral pathways, HBoV1 continues infecting toward the intestinal tract causing AGE. HBoV2 can be a causative agent of AGE and ARI in younger Amazonian children.

Predominance of Human Bocavirus Genotype 1 and 3 in Outpatient Children with Diarrhea from Rural Communities in South Africa, 2017-2018.

Human bocavirus (HBoV) is an emerging virus globally associated with diarrhea in young children. This study aims to investigate the prevalence of HBoV genotypes in children (≤5 years) from rural communities in South Africa (SA) suffering from acute gastroenteritis (AGE). A total of 141 fecal samples of children ≤5 years with acute gastroenteritis (AGE) were collected from rural primary health care facilities in the Vhembe district of SA between June 2017 and July 2018. Clinical symptoms and demographic data were also recorded. A total of 102 (72%) were outpatients, and 39 (28%) were hospitalized patients. Human bocavirus (HBoV) genotypes were determined using real-time multiplex PCR. DNA extracts of positive samples were confirmed by conventional PCR targeting the NS1 gene. Co-infection with other enteric viruses were determined in HBoV-positive samples using real-time PCR. HBoV was detected in eight (5.7%) children with AGE, of which three (37.5%) were HBoV1, three (37.5%) were HBoV3, and two (25%) were HBoV2. The majority of positive cases were identified in outpatients (62%) between the ages of 1 and 24 months. Co-infection in HBoV-positive samples with other enteric viruses included rotavirus (37.5%), adenovirus (37.5%), norovirus (25%), and astrovirus (12.5%). HBoV infections could be seen as a potential emerging diarrheal pathogen in South Africa. However, more studies are needed to understand the role of HBoV infections in children with AGE.

Prevalence of rotavirus and human bocavirus in immunosuppressed individuals after renal transplantation in the Northern Region of Brazil.

Immunosuppressive therapy causes severe impairment of host defense and diarrhea is a frequent complication in renal transplant recipients. This study aimed to describe the occurrence of Rotavirus A (RVA) and Human Bocavirus (HBoV) in fecal samples of immunosuppressed patients submitted to renal transplantation during posttransplant follow-up. A longitudinal study was carried out involving a 25-patient cohort, selected for kidney transplantation. A total of 126 fecal samples were collected between May 2014 and May 2016. Molecular techniques were used to detect and characterize circulating RVA and HBoV genotypes and statistical analysis were applied to verify the association between epidemiological and clinical characteristics. The prevalence of RVA and HBoV was 24% (6/25) and 40% (10/25), respectively. Among RVA and HBoV positive cases, the majority was female; did not conduct water treatment nor had adequate sewage facilities. The most detected genotypes were RVA G3 (62.5%) and HBoV-3 (95%). Phylogenetic analysis of HBoV strains indicated that studied samples were similar to those found in Asian and American countries. The present study point out the circulation of these viral agents among immunosuppressed individuals and these findings will enable the construction of new knowledge and care perspectives on the cause of diarrhea in this population.

Changing pattern of genotypic circulation of human bocavirus variants associated with acute gastroenteritis in Pune, Western India: A 5-year retrospective study.

Human bocavirus (HBoV) has been frequently associated with acute gastroenteritis. A 5-year retrospective study was undertaken to understand the circulation pattern and genotype distribution of HBoV in acute gastroenteritis cases in Pune, Western India. A total of 985 stool samples collected from sporadic acute gastroenteritis cases and asymptomatic controls (2007-2011) from children (≤5 years) were examined for the presence of HBoV. HBoV1 was detected throughout the study period while HBoV2 during 2007-2010, HBoV3 in 2007-2009 and in 2011, and HBoV4 in 2009 and 2011. Interestingly, HBoV2 was observed to be predominant in 2007 while HBoV1 and HBoV2 were detected at an equal frequency in the year 2008. HBoV1 was predominant from 2009 onwards. The present study highlights the changing pattern of genotypic circulation, emergence, and re-emergence of HBoV variants in acute gastroenteritis cases over a 5-year study period in Western India. The severity of gastroenteritis is reported to vary with HBoV genotypes. Thus, the present study emphasizes the need for routine HBoV surveillance in acute gastroenteritis cases from other parts of the country. This data will be valuable in the current scenario because implementation of rotavirus vaccination has led to the rising of other enteric viruses associated with the disease.

Molecular epidemiology of human bocavirus in children with acute gastroenteritis from North Region of Brazil.

Human bocavirus (HBoV) is a DNA virus that is mostly associated with respiratory infections. However, because it has been found in stool samples, it has been suggested that it may be a causative agent for human enteric conditions. This underpins the continuous search for HBoVs, especially after the introduction of the rotavirus vaccine due to acute gastroenteritis cases related to emergent viruses, as HBoVs are more likely to be found in this post-vaccine scenario. Therefore, the aim of this study is to demonstrate the prevalence of HBoV in children aged less than 10 years with acute gastroenteritis in Brazil from November 2011 to November 2012. Stool samples from hospitalized children ≤10 years old who presented symptoms of acute gastroenteritis were analysed for the presence of rotavirus A (RVA) by an enzyme-linked immunosorbent assay (ELISA), and for HBoV DNA by nested PCR. HBoV positivity was detected in 24.0 % (54/225) of samples. Two peaks of HBoV detection were observed in November 2011 and from July to September 2012. Co-infections between HBoV and rotavirus A were identified in 50.0 % (27/54) of specimens. Phylogenetic analysis identified the presence of HBoV-1 (94.8 %), HBoV-2 (2.6 %) and HBoV-3 (2.6 %) species, with only minor variations among them. Our findings provide evidence for the circulation of most HBoV genotypes (except HBoV-4) in the North Region of Brazil at a considerable rate and further investigations are necessary to improve our knowledge in the context of HBoV infections and their role in gastrointestinal diseases.

Human bocavirus detection and quantification in fecal and serum specimens from recipients of allogeneic hematopoietic stem cell transplantation: A longitudinal study.

The aim of this study was to evaluate the occurrence of human bocavirus (HBoV) and to determine viral loads in samples of patients admitted for allogeneic hematopoietic stem cell transplantation (allo-HSCT). Fecal and serum samples were collected from 19 patients, during a 24-month period. Samples were screened by quantitative polymerase chain reaction TaqMan assay, with specific probe and primers targeting the NP1 gene of all HBoVs genotypes (HBoV-1 to - 4), and viral loads were determined using serial dilutions of a recombinant plasmid. HBoV DNA was detected in 42.1% (8 of 19) of the patients in at least one type of sample (feces and/or serum) during the study period, with 75% (6 of 8) of the patients being positive in both types of sample. Viral shedding in feces had a median of 26 days (range, 5 to 121) and viremia was detected in 87.5% (7 of 8) of the patients. The HBoV loads in fecal samples were higher than in sera and, in most cases, HBoV was detected earlier in fecal than in sera samples. In six HBoV-positive patients (6 of 8) diarrhea was observed concomitantly to viral detection in fecal samples. A high frequency and loads of HBoV in allo-HSCT recipients was observed, especially in fecal samples. Positivity in fecal samples was an early predictor of HBoV presence.

Genomic recombination in primate bocavirus: inconsistency and alternative interpretations.

Human bocavirus (HBoV) is a single-stranded DNA virus in Parvoviridae family, causing respiratory diseases in human. The recent identifications of genomic recombination among the four human bocavirus genotypes and related non-human primate bocaviruses have shed lights into the evolutionary processes underpinning the diversity of primate bocavirus. Among these reports, however, we found inconsistency and possible alternative interpretations of the recombination events. In this study, these recombination events were reviewed, and the related genome sequences were re-analysed, aiming to inform the research community of bocavirus with more consistent knowledge and comprehensive interpretations on the recombination history of primate bocavirus.

Relative Abundance of Human Bocaviruses in Urban Sewage in Greater Cairo, Egypt.

Human bocavirus (HBoV) is predominantly found in the respiratory tract infections and in the stool of patients with gastroenteritis symptoms. However, data on the prevalence of HBoV genotypes in environmental samples are limited. Here we addressed the prevalence of HBoV in sewage collected from three different wastewater treatment plants in Egypt. HBoV-1, HBoV-2, and HBoV-3 were detected, whereas none of the samples were positive for HBoV-4. The median concentration of HBoV in influent samples was 8.5×103 GC/l for HBoV-1, 3.0×104 GC/l for HBoV-2, and 2.5×104 GC/l for HBoV-3. The concentration was reduced but not completely removed in the effluent samples. The median concentration in the outlet samples was 2.9×103 GC/l for HBoV-1, 4.1×103 GC/l for HBoV-2, and 2.1×103 GC/l for HBoV-3. Moreover, no seasonality pattern of HBoVs was found. The high incidence of HBoV in sewage samples provided an evidence of its circulation in the local population. Although the role of HBoV in respiratory or gastro-intestinal infections still remains to be fully elucidated, the risk of infection via contaminated water should be taken into consideration.

Molecular epidemiology and clinical severity of Human Bocavirus (HBoV) 1-4 in children with acute gastroenteritis from Pune, Western India.

Although acute gastroenteritis is a major public health problem worldwide, ∼40% of the cases remain undiagnosed for any etiological agent. Human Bocavirus (HBoV) has been detected frequently in feces of diarrhoeic children suggesting its possible etiological involvement in the disease. HBoV has not been reported in association with acute gastroenteritis from India. Fecal samples (n = 418) collected from children (age ≤5 years) hospitalized with acute gastroenteritis, between January 2009 and December 2011, from three local hospitals were examined for presence of HBoV using PCR targeting the partial VP1/VP2 capsid region (∼575 bp) followed by phylogenetic analysis. HBoV was detected in 24/418 (5.7%) cases. Co-infection was observed in 5/24 (21%) cases. HBoV infections occurred in children ≤12 months of age. Peak HBoV activity was observed in monsoon and post monsoon season. All four HBoV genotypes were detected in the study region. Major clinical symptoms of HBoV mono infections included diarrhoea (100%), fever (90%), dehydration (74%), and vomiting (58%). Dehydration was observed in all of the HBoV2-4 cases and in 50% of the HBoV1 cases. Clinical severity varied with genotype (HBoV2 > HBoV1 > HBoV3 > HBoV4). HBoV2 cases recorded severe and very severe infections. The study illustrates prevalence and vast genetic diversity of HBoVs in acute gastroenteritis. It highlights the clinical features of HBoV1-4 infections and sheds light on clinical impact of HBoV genotypes in gastroenteritis. J. Med. Virol. 89:17-23, 2017. © 2016 Wiley Periodicals, Inc.

Human bocavirus: Current knowledge and future challenges

Human bocavirus (HBoV) is a parvovirus isolated about a decade ago and found worldwide in both respiratory samples, mainly from early life and children of 6-24 mo of age with acute respiratory infection, and in stool samples, from patients with gastroenteritis. Since then, other viruses related to the first HBoV isolate (HBoV1), namely HBoV2, HBoV3 and HBoV4, have been detected principally in human faeces. HBoVs are small non-enveloped single-stranded DNA viruses of about 5300 nucleotides, consisting of three open reading frames encoding the first two the non-structural protein 1 (NS1) and nuclear phosphoprotein (NP1) and the third the viral capsid proteins 1 and 2 (VP1 and VP2). HBoV pathogenicity remains to be fully clarified mainly due to the lack of animal models for the difficulties in replicating the virus in in vitro cell cultures, and the fact that HBoV infection is frequently accompanied by at least another viral and/or bacterial respiratory and/or gastroenteric pathogen infection. Current diagnostic methods to support HBoV detection include polymerase chain reaction, real-time PCR, enzyme-linked immunosorbent assay and enzyme immunoassay using recombinant VP2 or virus-like particle capsid proteins, although sequence-independent amplification techniques combined with next-generation sequencing platforms promise rapid and simultaneous detection of the pathogens in the future. This review presents the current knowledge on HBoV genotypes with emphasis on taxonomy, phylogenetic relationship and genomic analysis, biology, epidemiology, pathogenesis and diagnostic methods. The emerging discussion on HBoVs as true pathogen or innocent bystander is also emphasized.

A novel primer set for improved direct gene sequencing of human bocavirus genotype-1 from clinical samples

Human bocavirus genotype (HBoV-1) is a parvovirus associated with respiratory tract infections in children with different degrees of severity. The current study intended to improve the direct gene sequencing of the HBoV-1 using a newly developed primer set. Screening the presence of human bocavirus infection among in-patients children suffering from lower respiratory tract infections was another aim of the current study. Nasopharyngeal swab samples from in-patients children suffering from lower respiratory tract infections were examined. The real-time polymerase chain reaction was used for the initial screening as a highly sensitive method to detect the HBoV. Genotyping of real-time positive samples was attempted by direct sequencing of PCR amplicons using NP, VP1/2 and the newly developed VP/NC primers. HBoV-1 was present in 56.8% of the examined children. The newly developed primer set successfully amplified all real-time PCR positive samples, however, the other primer pairs did not reliably detect real-time PCR positive samples. The gene sequences of the detected HBoV-1 showed conserved sequences to each other with a low rate of discrepancies. The high rate of infection and the similarity between the detected strains strongly suggest nosocomial infections.

Detection of human bocavirus type 1 infection in Panamanian children with respiratory illness.

The human bocavirus (HBoV) was added as a new member of the Parvoviridae family in 2005 upon its discovery in nasopharyngeal aspirates from children with respiratory infection. Recently, there has been increasing evidence of worldwide circulation of HBoV; however, in Latin America few studies have been conducted. In order to detect the circulation of HBoV in Panama, based on the National Flu Surveillance System, we developed this retrospective, cross-sectional study, from January 2011 to January 2012. Children younger than 6 years old who presented with respiratory disease were enrolled in this study. Nasopharyngeal swabs were taken in sentinel surveillance sites. Samples were tested to detect mRNA from HBoV, as well as viral RNA and DNA from others respiratory viruses. A total of 1078 patients were enrolled in this study. Overall, 44 (4.1%) of the patients presented HBoV. The most common symptoms were cough (84.6%), fever (82.1%), rhinorrhea (74.4%), and sore throat (38.5%). Less than half (45.5%) of HBoV infected patients presented with monoinfection while 54.5% of cases presented with coinfection with others respiratory viruses. Both, outpatients and inpatients were included in this study. Outpatients corresponded to 52.3% of the cases and 47.7% were inpatients. Coinfection was observed in the 50% of the inpatient cases. Phylogenetic analyses indicated that the circulating strains belonged to different clades of HBoV genotype 1. Taken together, our results support the pathogenic nature of this viral agent, especially in younger children.

Prevalence analysis of different human bocavirus genotypes in pediatric patients revealed intra-genotype recombination

BackgroundHuman bocavirus (HBoV) genotypes 1–4 have been detected worldwide in respiratory samples and stool samples, and are increasingly associated with respiratory and intestinal infections of previously unknown etiology in young children. Several studies revealed evidence of extensive recombination among HBoV genotypes at the NP1 and VP1 gene boundary region. This study explored the prevalence of HBoV genotypes in pediatric patients in Beijing, and studied their phylogeny. MethodsA total of 4941 respiratory specimens and 1121 fecal specimens were collected from pediatric patients with respiratory infections from January 2006 to December 2013, or with acute diarrhea from October 2010 to December 2012. Conventional PCR was used to detect HBoV1–4 within these samples. Gene fragments at the NP1 and VP1 gene boundary were amplified from HBoV-positive specimens, sequenced, and their phylogenetic inferences constructed using MEGA 6.0 software. Recombination events were identified with SimPlot software. ResultsHuman bocavirus 1, 2, and 3 were detected in 9 (0.80%), 15 (1.33%), and 1 (0.08%) of 1121 stool samples, respectively. However, only HBoV1 (82, 1.65%) was detected in respiratory specimens. Phylogenetic analysis of gene fragments at the HBoV NP1 and VP1 gene boundary indicated that HBoV1 sequences obtained from fecal or respiratory specimens across 8years were highly conserved (99–100%), while 15 HBoV2 sequences collected across 2years in Beijing were more diverse with up to 4.40% variation. Of the 15 HBoV2 sequences, 14 clustered into a new lineage divergent from other HBoV2 sequences in GenBank. Five HBoV2 genomic sequences were analyzed for recombination, revealing intra-genotype recombination between HBoV2A and HBoV2B. ConclusionsMore HBoV1 were detected in children with respiratory tract diseases, and HBoV2 in patients with acute diarrhea. Phylogenetic analysis revealed a new cluster of HBoV2 was prevalent in China, which may be the result of intra-genotype recombination between HBoV2A and HBoV2B.

The genomic and seroprevalence of human bocavirus in healthy Chinese plasma donors and plasma derivatives.

Human bocavirus (HBoV) is a novel parvovirus identified in 2005. It has mostly been detected in respiratory and enteric infections and has not been studied large scale in blood products in relation to transfusion. An in-house quantitative polymerase chain reaction (Q-PCR) was developed to test HBoV DNA in plasma and plasma derivatives. Plasma samples (n = 6096) collected from healthy donors, 241 plasma pools, and 326 plasma derivatives were screened for HBoV DNA by Q-PCR. Positive samples were confirmed by nested PCR and further amplified for sequence analysis and phylogenetic studies. The prevalence of immunoglobulin (Ig)G and IgM specific to HBoV structural proteins was measured by enzyme-linked immunosorbent assay in 209 samples grouped according to virus load (Group 1, HBoV DNA >10(4) copies/mL; Group 2, HBoV DNA >5 × 10(2) copies/mL but below 10(4) copies/mL; Group 3,HBoV DNA negative). The genomic prevalence of HBoV in the plasma donors was 9.06%, ranging from 5.01 × 10(2) to 3.02 × 10(6) copies/mL. HBoV-specific IgG and IgM were detected at 20.00 and 7.50% in Group 1, at 20.29 and 2.90% in Group 2, and at 13.00 and 4.0% in Group 3, respectively. Phylogenetic analyses proved that HBoV Genotype 1 was the prevalent genotype in Chinese plasma donors. Low levels of HBoV DNA were detectable at high prevalence in Chinese plasma donors and plasma derivatives. Further study is needed to determine whether HBoV screening is necessary.

Evolutionary time-scale of primate bocaviruses

Human bocavirus (HBoV) is associated with acute gastroenteritis in humans, occurring mostly in young children and elderly people. Four bocavirus genotypes (HBoV1–HBoV4) have been found so far. Since there were no data on the contribution of HBoV to gastroenteritis in Russia, 1781 fecal samples collected from infants hospitalized with acute gastroenteritis in Novosibirsk, Russia during one year were tested for the presence of nucleic acids from HBoV and three major gastrointestinal viruses (rotavirus A, norovirus II, and astrovirus). HBoV was detected only in 1.9% of the samples: HBoV1 was detected in 0.6% and HBoV2, in 1.3%. Complete genome sequencing of three Novosibirsk isolates was performed. An evolutionary analysis of these sequences and the available sequences of human and great apes bocaviruses demonstrated that the current HBoV genotypes diverged comparatively recently, about 60–300years ago. The independent evolution of bocaviruses from chimpanzees and gorillas commenced at the same time period. This suggests that these isolates of great apes bocaviruses belong to separate genotypes within the species of human bocavirus, which is actually the primate bocavirus. The rate of mutation accumulation in the genome of primate bocaviruses has been estimated as approximately 9×10–4substitutions/site/year. It has been demonstrated that HBoV1 diverged from the ancestor common with chimpanzee bocavirus approximately 60–80years ago, while HBoV4 separated from great apes bocaviruses about 200–300years ago. The hypothesis postulating independent evolution of HBoV1 and HBoV4 genotypes from primate bocaviruses has been proposed.