Posted on Authorea 7 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159414162.24879023 — This a preprint and has not been peer reviewed. Data may be preliminary. ewe .%ad6%(age l,21;Zage l,21;Zo ta. 08.C-neto fPBoV of Co-infection 2018). al., et Zhou 2015; al., prevalence et the with Zhang China 2014; in regions al., or et provinces (Wang 20 2014). in 64% al., reported and been et into have 7.3% (Zhou classified PBoV between G3 be with PBoV infected to and pigs proposed date, G2, was To PBoV PBoV G1, And PBoV PBoV-7V. groups, and different 1( PBoV-6V ( three phosphoprotein PBoV3C, 1 nuclear PBoV5, encodes protein PBoV4, ORF3 nonstructural PBoV3, 2009), ( a al., (ORFs), 2 encodes et frames the and ORF1 (Sun reading region 1 2009). open N-terminal proteins three al., al., additional caspid of et et consists viral (Arthur (Zhou which Africa ORF3 encodes virus and and DNA America ORF2, single-stranded et North ORF1, non-enveloped (Zhai Asia, a 2010 Europe, is in in PBoV PBoV1 identified or been PBo-likeV (PBoV) with has bocavirus pigs PBoV 2009). porcine of then, al., 2014). as nodes Since lymph et named from and (Blomstrom 2010). reported China Sweden al., first in in was discovered bovine Li (PBo-likeV) (PMWS) subsequently (CMV), 2008; virus was syndrome al., virus boca-like wasting et minute porcine (Lau multi-systemic (CslBoV) 2009, canine post-weaning bocavirus In (PBoV), lion family sea 2011). bocavirus the California al., and porcine et to (GBoV), (HBoV), bocavirus belongs gorillas bocavirus (BPV), which human bocavirus genus includes classified and novel , a is Bocavirus molecular 1 the of understanding our extend they results These and PBoV-TY. China, PBoV. and central of CH/HNZM in evolution for prevalence ZJD and found PBoV strain was epidemiology the PBoVs reference of strains of information with PBoV acids further Compared from amino provided derived NS1 had analysis. the be may in recombination variation which the higher (KJ755666) through a strain (HM053694)-China, GD18 (KF206160) PBoV2 strain 57AT-HU the PBoV and into pairwise the clustered (MH558677) reference to PBoV-TY the 35 0912/2012 related and with that CH/HNZM closely identity strains was showed sequence PBoV PBoV-TY nucleotide alignment two and genomic PBoV-TY the sequence group, 44.5%-95.8% that and genomic demonstrated had analysis they multiple KX017193) phylogenetic and PBoV3/4/5. The A 94.8%, numbers was and strains. study. strains (accession PBoV1/2 PBoV this two CH/HNZM both the positive in for strains of were cloned positive similarity China, (52.67%) PBoV were were central 148 of MH454686) (19.57%) samples, of numbers 55 sequences 281 farms and (accession the genome pig 236 PBoV3/4/5, Among different complete for including 27 PBoVs. two positive detect samples in were Subsequently, to piglets clinical (41.63%) developed diarrheal 117 281 were from PBoV1/2, assays (PBoV), collected for qPCR were bocavirus I-based samples Green porcine fecal SYBR of 45 and analysis and evolutionary samples tissue and intestinal phylogenetic the investigate To Abstract 2020 7, July 1 Chen Zheng Lan-Lan China central in of bocavirus analysis porcine evolutionary and phylogenetic detection, Molecular ffiito o available not Affiliation INTRODUCTION VP 1 and 1 VP eune,Po a encasfidit ieetcae,wihicue BV,PBoV2, PBoV1, includes which clades, different into classified been has PBoV sequences, 2 1 inToCui Jian-Tao , 1 a Qiao Han , VP1/2 ,and ), 1 i-hn Li Xin-Sheng , 1 VP1 ossso h entire the of consists Parvoviridae 1 ioagLi Xiaokang , subfamily , VP2 1 n Hongying and , NP1 euneadan and sequence NS1 Parvovirinae .Bsdon Based ). ,ORF2 ), Posted on Authorea 7 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159414162.24879023 — This a preprint and has not been peer reviewed. Data may be preliminary. C ecin eeotmzd n h estvt,seict n erdcblt eeevaluated. were reproducibility real-time and I-based specificity Green sensitivity, SYBR the the and of Tris–HCl, parameters optimized, (10mM The were buffer respectively. reactions generated triplicate, PCR TE were in PBoV3/4/5 in 1 performed and (from diluted and PBoV1/2 number pMD18-209, of serial copy detection 10-fold known for curves and a 260 standard with determined at USA). PCR MA, were absorbance real-time Waltham, The ultraviolet plasmids Scientific, EDTA). Fisher by 1mM of (Thermo quantified numbers micro-spectrophotometer were Nano-100 copy plasmids a The using standard nm two 280 the and nm into of transformed purities then and and concentrations respectively, recombinant pMD18-209 to The and according INC) China). pMD18-327 coli Dalian, as Biotechnologies. (Takara, (Watson vector designed PBoV3/4/5, and pMD18-T Kit were and into Mini PBoV1/2-F/R plasmids cloned Extraction PBoV1/2 and primers Gel positive instructions, of the with The manufacturer’s USA). using the detection Research, amplifications purified (MJ were for cycler PCR thermal products Conventional Peltier assays PCR PCT-200 the PCR using generated. performed real-time were were PBoV3/4/5-F/R I-based curves Green standard SYBR the the assays PCR develop real-time To GreenIbased SYBR the of to Development according covering 2.3 fragments addition, designed overlapping (Table.1). In were three genome 1). Q3F/Q3R) amplify (Table viral and to entire bp HM053693) Q2F/Q2R the number 209 (Q1F/Q1R, accession of primers (GenBank fragment sequence PCR a reference specific amplifying of JF429834, num- pairs number the accession three accession targeting (GenBank GenBank PBoV3/4/5-F/R JN831651), PBoV3/4/5 of and primers of sequence JN621325 of sequences primers KF206158, pair nine JF429836, specific a JN681175, The of GenBank and JF713715, 1). alignment of KC473563, (Table an sequence JF713714, bp the JF429834, 327 from on ber of found based product were PBoV1/2- software amplification PBoV3/4/5 primers USA) an 6.0, of for gave (version which pair Premier HM053693, a Primer number and the accession DNASTAR KM402139), using and designed 7.1, was HQ291309, KF206161 to (version HM053694, F/R KF206157, HM053693, software the number subjected KF206155, accession within KF025393, DNAStar and (GenBank sequences sequences KF025392, of GenBank genomic conserved PBoV1/2 program the nine highly of from MegAlign The gnment downloaded USA). the Madison, were using Inc., PBoVs analysis of homology sequences perform nucleotide -80 genome at Eighteen stored and according calculated China) was design Ltd., DNA Primers Co, of at 2.2 Shanghai concentration centrifuged Biotech The (Sangon and diluted instructions. Kit were manufacturer’s vortexed Isolation Samples the were DNA to China. suspensions Genomic were central samples Virus the of fecal Column and cities 45 UNlQ-10 18 pH7.4), and in samples (PBS, 000 farms tissue saline 12 pig intestinal phosphate-buffered different 236 27 including with in samples 1:10 piglets 281 of diarrheal total from a collected 2019, to 2016 preparation From DNA and PBoV collection the Sample investigate China. 2.1 central to from the aimed However, PBoV METHODS In we of 2010). AND properties study, al., MATERIALS sequence 2015). current et genome 2 co- al., the (Zhai full-length the piglets analyze In et and in and Luo pigs diarrhea recognized. pigs, suggesting diarrheal 2014; causing further pigs, in healthy healthy in prevalence al., needs of in role that et PBoV than important than pigs of Zhou an pigs diarrheal play pathogenicity of 2014; diseased might samples in al., PBoV in higher the rate et was that PBoV infection 2010; Huang and al., higher PEDV 2014; of et significantly rate (Blomstrom al., infection a (CSFV) et has virus virus Zhang fever PBoV syndrome swine 2013; addition, respiratory porcine classic (PEDV), and al., and virus reproductive et (PTTV) diarrhea virus porcine McMenamy epidemic (PRV), teno porcine torque virus as porcine pseudorabies such (PRRSV), (PCV2), reported, been 2 also type has circovirus viruses porcine other with DH-5 × α o i t4 at min 5 for g optn el TKR) oiiecoe eeioae n umte o eunig The sequencing. for submitted and isolated were clones Positive (TaKaRa). cells competent ° oclettecaie uentns h ia N a xrce sn an using extracted was DNA viral The supernatants. clarified the collect to C × 10 9 o1 to × 10 0 copies/ 2 VP μ eeo BV// a eindbsdo the on based designed was PBoV3/4/5 of gene 1 )o h eobnn lsispD837and pMD18-327 plasmids recombinant the of L) NS1 eewr eemndfo nali- an from determined were gene ° Escherichia C. Posted on Authorea 7 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159414162.24879023 — This a preprint and has not been peer reviewed. Data may be preliminary. o h w rmrst QFQRadQFQR,PRpooo a are u n25 in out 94 carried at was protocol PCR 0.5 Q2F/Q2R), and (Q1F/Q1R 12.5 sets mixture: primer two the For amplification sequence genome 20 Complete a 2.4 95 in of performed cycles was 39 PCR by followed Australia). 6 using Research, and performed (Corbett primer were cycler PBoV3/4/5 thermal and real-time PBoV1/2 μ 2000 of amplifications Gene PCR Rotor real-time the I-based Green SYBR The t72 at a 47%i eta hn 2021,adi a uhhge hnta fpeiu eot Zaget (Zhang reports previous of that than higher much was 55 it PBoV and and of PBoV3/4/5, prevalence (210/281), the for China Overall, positive PBoV3/4/5. central (148/281, were and in samples PBoV1/2 41.63%) 148 both 74.73% (117/281, for samples, was samples positive clinical were 281 117 19.57%) of PBoV1/2, (55/281, results samples for detection positive PCR specific, were I-based sensitive, Green 52.67%) highly ranged SYBR with CV the I-based use to Green and to SYBR According SD good the inter-assay really that showed were the results detection of These PBoV3/4/5 reproducibility. values respectively. and and 2.304%, The PBoV1/2 intra-assay to respectively. for % and 0.678%, assays 0.768 ranged inter- qPCR and to (CV) variation 0.539 using 0.076% of to triplicate and co-efficient 0.112 and in from 0.042 (SD) determined to deviation was standard 0.010 intra-assay assays from the the 88 of values was of The (Tm) reproducibility comparisons. temperatures The -3.6726x melting peaks. = the 85 melting y that of peak was copies/ Tm melting 33 PBoV1/2 the was specific of limit a detection curve with minimum detected standard the and The 0.99, samples China. of clinical central in of PBoV3/4/5 R copies/ and cities with PBoV1/2 18 detect 41.425, in to + piglets developed 0.01. were diarrheal was assays value from four qPCR P I-based et acceptable than Green (Zhai highest more the SYBR previously by bp, described 20 DISCUSSION as supported to AND method events set RESULTS detection was recombination 3 single size those window a the on Only and dependence PBoV. 2010), avoid events al., of to recombinant detected deemed genomes to were complete used programs were two 4.39) (RDP the and program SiScan in detection Chimaera, Recombination MaxChi, in BOOTSCAN, embedded GENECONV, replicates. 3Seq) (RDP, using 1,000 methods of detection performed recombination bootstrap (version was Seven software a analysis (MEGA) and Phylogenetic Analysis distances, program Genetics USA). MegAlign Kimura Evolutionary the with Inc., Molecular using 6.0) DNASTAR the analyzed in 7.1, and method aligned (version minimum-evolution were software the 2) using DNAstar (Table similarities strains the sequence reference for of of 35 screened sequences and were genomic strains The study PBoV GenBank. this two in in database the obtained against strains (http://www.ncbi.nlm.nig.gov/BLAST) BLAST PBoV two of sequences sequencing analyses Genomic all phylogenetic and and sequencing, alignment for Sequence (TaKaRa) 2.5 Vector pMD18-T duplicate. in the into performed were cloned reactions were products purified The 60 at 95 at pre-denaturation follows: 25 in out carried 3 was protocol PCR sets, μ ouecnann 3 containing volume L fsml N,0.5 DNA, sample of L μ fec rmr(50 primer each of L ° ° ° o i;floe y3 ylso eauaina 98 at denaturation of cycles 35 by followed min; 3 for C o 5,etnina 72 at extension 45s, for C o mn n hnfloe yafia xeso tpa 72 at step extension final a by followed then and 2min, for C μ .Tesadr uv fPo // a eemndt ey=-.39 917 ihR with 39.137, + -3.3379x = y be to determined was 3/4/5 PBoV of curve standard The L. ° μ μ .Hwvr rnmsil atoneii iu TE) EV RS n R a no had PRV and PRRSV PEDV, (TGEV), virus gastroenteritis transmissible However, C. f2hnaMxBffr 2 Buffer, Max 2Phanta of L itle ae.Teapicto odtoswr 95 were conditions amplification The water. distilled L 2 aus(qaeo h orlto offiin)o .9 n h eeto ii a 16.6 was limit detection the and 0.99, of coefficient) correlation the of (square values μ N fsmls 10 samples, of DNA L μ μ ° fec rmr(50 primer each of L o 0s 58 s, 30 for C ) n 8 and M), ° ° o mn olwdb 5cce fdntrto t95 at denaturation of cycles 35 by followed 5min; for C o mn n hnfloe yafia xeso tpa 72 at step extension final a by followed then and 2min, for C μ fddH of L μ ° o 5sad72 and s 25 for C μ fterato itr:12.5 mixture: reaction the of L fsml N,1 DNA, sample of L 2 μ .TePRprmtr eea olw:pre-denaturation follows: as were parameters PCR The O. f2 of L μ ) n 8.5 and M), 3 × YRPei Ex Premix SYBR μ .Frteeauto fseict,Po12was PBoV1/2 specificity, of evaluation the For L. ° o 0s. 30 for C ° o 0s neln t55 at annealing s, 10 for C μ μ fPat a ue,0.5 Super, Max Phanta of L fddH of L ° o 0mn o h 3/3 primer Q3F/Q3R the For min. 10 for C ° o i siiildenaturation initial as min 3 for C 2 μ .TePRprmtr eeas were parameters PCR The O. Taq f2sTqMse i (Dye), Mix Master Taq 2Es of L ,0.5 I, ° n BV// with PBoV3/4/5 and C μ (25 L ° ° o 5s extension s, 15 for C o 5,annealing 45s, for C μ ftereaction the of L μ μ fdT Mix, dNTP of L o/)o each of mol/L) ° o 0min. 10 for C 2 values Posted on Authorea 7 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159414162.24879023 — This a preprint and has not been peer reviewed. Data may be preliminary. thsbe lutae ocnancnevdmtf soitdwt oln-icerpiain eiaeand helicase replication, rolling-circle with associated the compared motifs After conserved 2011). al., contain et to (Lau activities illustrated ATPase been in samples has porcine of It acids in amino time encoding the first and that comparison, ORF1 the the for as strain identified this that chose We nt (HM053694)-China 2010). 5186 al., ZJD VP2 et strain with (Cheng PBoV sequence et study, 2010 in this genome (Kapoor China in full-length parvoviruses Minor listed nearly we of tropism. strains a range reference is tissue host PBoV 28 and of and the acids potential Among immunogenicity amino pathogenic analyzed. the 2013). study, with the this swine. al., alter In relationship of et 2012). could PBoVs al., Zhang close proteins reemerging 2011; have of and al., changes parvoviruses emerging et genetic the HBoVs, of (Malecki to to origin proteins attention related zoonotic more Capsid closely of pay are to be vital strains could tree is PBoV HBoV phylogenetic it that some of Thus, Moreover, hypothesis result the the sequences. On to to nucleotide identical BPV1. leading genomic was and which the HBoV PBoV2, of with to altogether analysis belonged cluster the PBoV-TY of and large analysis CH/HNZM a tree strains in phylogenetic was the the of their PBoV3/4/5 on basis on whilst the Based based were strains, bocaviruses. cluster CMV strains other large and 7 PBoV3/4/5 with a CslBoV1 whilst branch in another strains, were in CMV situated strains VP1 were and PBoV2 strains CslBoV1 and PBoV3/4/5 with whilst PBoV1 (Fig.S2), altogether branch. on independent cluster based trees closer an large were phylogenetic in a strains the PBoV2 in Additionally, and placed HBoV. PBoV1 were that of of showed which source results sequences groups, The further PBoV2 acid structed. or a amino PBoV1 the be with on might that based group than PBoV3/4/5 HBoV Interestingly, PBoV3/4/5 with (Fig.1). that relationship (44.5%-55.3%) with indicated phylogenetic groups branch closer phylogene- HBoV large were a or they showed a However, these PBoV3/4/5 group formed homology. PBoV1, and sequence group, two nucleotide from 94.8%, PBoV2 The 86.8%-95.8% distinct the was PBoV3/4/5. had tically into and and strains strains, clustered PBoV2 PBoV PBoV2 were PBoV1, 12 two PBoV-TY clusters, other the and three into of CH/HNZM nucleotide divided strains similarity genomic be PBoV on pairwise could based the strains tree that PBoV phylogenetic representative showed the constructing results by The analyzed nt sequences. were 2,112 2) CH/HNZM with (Table strain length, strains in reference PBoV nt of 5156 length comprised genomic / PBoV-TY The strain alignment PBoV ORFs. the The putative then respectively. and three encoding acquired had nt, were 5173 also PBoV-TY) was they and that (CH/HNZM showed strains PBoV. However, PBoV PBoV analyses to (47.83%). found that of pigs associated 2015) genomes healthy reported disease al., complete with clinical 2016) et Two compared definite (Zhang al., when no al. (73.95%) et has et pigs (Pfankuche 2012; there diarrheic Zhang al. Meng, encephalomyelitis. now, in until PBoV triggers 2010; et of that al., Pfankuche incidence pathogen et 2015). higher a (Blomstrom a al., as CSFV co-infection et considered the PRRSV, addition, Luo be as In should such 2013; PMWS. PBoV common, of al., higher development was et the the pathogens in McMenamy et that other role (Blomstrom demonstrated with previously a study play reported PBoV might results this samples of to PBoV in positive similar that were the PBoV suggested which and all and 2010), PWMS, (216/281), in al., PCV2 with found 76.87% pigs of was was from rate PCV2 samples PEDV in co-infection (138/281). of prevalence 49.11% high rate pigs. was The infection PEDV in PBoV. The and diarrhea detected. centralof PBoV of also of in cause south were rate piglets probable co-infection PCV2 and diarrheal the a However, and east in frequency. PEDV as study, the farms thought occurrence this swine in be PBoV In among should distributed high virus distributed mainly of this broadly center PBoV that has the that indicating PBoV was showed China, that province which suggested Jiangsu China results and in our China, 2018b) of al. areas et coastal Zhou 2015; al. VP2 and and ee(,0 nt (1,704 gene NP1 VP2 ftetoPo tan eecmae ihterfrnesri J (HM053694)-China. ZJD strain reference the with compared were strains PBoV two the of mn cdsqecs(i.3 4,Po1adPo2fre ag lse loehrwith altogether cluster large a formed PBoV2 and PBoV1 S4), (Fig.S3, sequences acid amino NS VP2 NS1 slctda h ’edo h BVgnm,adi seta o N replication. DNA for essential is and genome, PBoV the of end 5’ the at located is 1 212nt), (2,112 ,ad60nt 690 and ), VP1 NS1 NP1 NS and , NP1 1, ee h eei eainhp ftoPo tan ih35 with strains PBoV two of relationships genetic The gene. NS1 P,VP2 VP1, VP2 , , VP1 4 215n,icuig171nt 1,701 including nt, (2,115 NP1 NS and , and VP1 mn cd ftetoPo tan ihthe with strains PBoV two the of acids amino 1 VP2 NP1 and rtisfral3 oaiue eecon- were bocaviruses 37 all for proteins ftePo tan eecmae and compared were strains PBoV the of VP2 rti eune,tetoPBoV two the sequences, protein NS1 VP2 NS1 nlss(i.1 and (Fig.S1) analysis and ) ee 18nt 2118 gene, NP1 NP1 NS sequences 67nt), (687 1, VP1, VP1 Posted on Authorea 7 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159414162.24879023 — This a preprint and has not been peer reviewed. Data may be preliminary. a 9.891 = Val h uhr elr htte aen oflc finterest. of number conflict STATEMENT AVAILABILITY no [grant DATA have they China that declare authors of The Foundation INTEREST Science OF Technology University CONFLICT Natural Agricultural Henan KJCX2018C02]. and National number le- 204200510015] [grant number high Fund [grant the Zhongyuan Innovation plan 2019GGJS051]; support number by special [grant talents Province vel supported Henan of Foundation was Teacher 31802218];Young work to submitted This been have respectively. PBoV-TY MH454686, and and KX017193 CH/HNZM ACKNOWLEDGEMENTS are strains numbers PBoV accession of and sequences GenBank, the genome information complete valuable two provide The study PBoV. the this of numbers and characteristics of accession biological China, results sequence and central The Nucleotide epidemiology China. in common. of prevalent central very analysis are in is further farms PBoVs the PCV2 pig that recombination for or in indicates and PEDV PBoV phylogenetic study with characterized, of present were prevalence co-infection The strains the conducted. field demonstrate were PBoV2 to of analyses study future. sequences PBoV1/2 first genome the with with complete in the infection 2013, Two recombination mixed is in of of this PBoV-TY prevalence possibility China conclusion, or high the In of a provided CH/HNZM Besides, which province strain bocavirus. reported, Guangdong PBoV genus was from the that PBoV3/4/5 clustered of and GD18 demonstrated was members results strain PBoV-TY novel strain The be PBoV identity. may PBoV with nucleotide the altogether PBoVs, genomic branch other 94.2% with small PBoV-TY a or in CH/HNZM strain PBoV either programs 7 by supported were which (KF206160), 3.158 recombination 3.85 57AT-HU = The was P-Val 2011). strain Av. al., parent (RDP, minor et located the Zeng strain to (KJ755666) and 2011; capacity GD18 (MH558677) al., in their existed improve et was which (Lau breakpoint the recombinant in worldwide PBoVs, and putatively Parvovirinae, one hosts of the that Suidae genotypes revealed of analysis among new evolution the spread of for and generation mechanism adapt the important increase an is could mutations recombination point ZJD PBoV-TY of strain and accumulation reference CH/HNZM The with strains PBoV comparison between when found sites were variation sites aa (Table.S1). variation four aa were two There and 2009). (HM053694)-China, al., et (Sun replication in the both PBoV-TY) contained and the PBoV-TY (CH/HNZM encodes in strains strain sites PBoV PBoV variation two aa and the three sites, contained variation PBoV-TY of aa strain in basis sites major the variation three On aa 2010). contained two al., USA CH/HNZM et the strain (Cheng in infectivity PBoV parvovirus pigs for from critical strains is VP2 that PBoV activity with (sPLA2) inconsistent A2 was unique proteins the which capsid the in the study, the in between variation codes was this ORF2 sites higher 2014). in variation al., a found aa et Moreover, (Jiang six was (Table.S1). were PBoVs PBoV-TY There of and sites, sites. variation acids variation CH/HNZM aa aa strains major four PBoV eight contained two contained PBoV-TY CH/HNZM strain strain PBoV PBoV and (HM053694)-China, ZJD strain reference Bocavirus × mn cd ftetoPo tan n oprsnwt eeec tanZD(HM053694)-China, ZJD strain reference with comparison and strains PBoV two the of acids amino VP1/2 10 -14 NP1 aCi v -a 1.89 = P-Val Av. MaxChi, ; × eea oiin348t ,8 to h eoe(i.) h ao aetsri a 0912/2012 was strain parent major The (Fig.2). genome the of nt 4,986 to 3,458 position at gene eu.Atog h ucinof function the Although genus. 10 rti hti oae ewe R1adOF,wihi hrceitcgntcfaueof feature genetic characteristic a is which ORF2, and ORF1 between located is that protein VP1 -19 Sq v -a 8.082 = P-Val Av. 3Seq, ; rti ( protein VP1 × 10 BVsri HHZ otie ormjra aito ie,adPBoV and sites, variation aa major four contained CH/HNZM strain PBoV . VP1 -14 EEOV v -a 5.595 = P-Val Av. GENECONV, ; )(age l,21) hsdmi niae ertr phospholipase secretory a indicates domain This 2012). al., et (Yang u) × 10 NP1 -12 × hmea v -a 1.988 = P-Val Av. Chimaera, ; VP1 10 nPo ean nla,i sesnilfrCVDNA CMV for essential is it unclear, remains PBoV in 5 -14 VP2 .Atog orcmiainsga a on for found was signal recombination no Although ). and hr eeol n avrainst between site variation aa one only were There . VP VP2 and 1 n osre YGF oi domain motif “YXGXF” conserved a and , VP × 10 mn cd TbeS) ORF3 (Table.S1). acids amino 2 -10 OTCN v -a = P-Val Av. BOOTSCAN, ; × 10 -13 icn v P- Av. SiScan, ; NS VP1 amino 1 and Posted on Authorea 7 Jul 2020 — The copyright holder is the author/funder. All rights reserved. No reuse without permission. — https://doi.org/10.22541/au.159414162.24879023 — This a preprint and has not been peer reviewed. Data may be preliminary. aek,M,Shlgn . cide,O 21) ua oaiu:silmr usin hnanswers. than questions more still bocavirus: I. Human W. (2011). Lipkin, O. https://10.2217/fvl.11.78 Schildgen, . 6:1107-1114. . & . Virology, Future V., L., Journal Schildgen, disease. 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A,Mxh,Ciar,SSa n Sq ( BOOTS- 3Seq) GENECONV, and (RDP, are SiScan programs numbers RDP4.39 Chimaera, by accession MaxChi, events CAN, GenBank recombination the PBoV potential in two and of Determination The circles, method Fig.2 replicates. solid neighbor-joining 000 black the 1, with from marked by parentheses. are node in generated each and study shown was for present CH/HNZM indicated the tree strains were from PBoV The values strains two strains. bootstrap of and reference sequences software, 35 genome MEGA7 complete with the together on PBoV-TY of based analysis analysis Genetic Phylogenetic (2018b). Fig.1 D. of A. Archives Qian, NS1. & in legends R., breakpoint Chen, Figure recombinant X., natural Z. a Lin, of https://10.1007/s00705-017-3606-8 F., identification 163:707-712. Q. 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