Laboratory and EpidemiOlogy Communications
The Isolation of Human Parechovirus 1 from Cases ofAcute ResplratOry Illness in Children
Shinichi Takao*, Shinji Fukuda, Yukie Shimazu, Masahiro Noda and Shizuyo Tokumoto
Division ofMicrobiology II, Hirosh ima Prefectural Institute of Health and Environment,
Minami‑machi I‑6‑29, Minami‑ku, Hiroshima 734‑0007
Communicated by Hiroo lnouye(Accepted April 3, 200 1 )Picornaviridae is a large family of single‑strand positive‑

infections caused by HPEVl.
From September 1999 to December 2000, a total of 2,257sense RNA virusesthatincludes a number of important humanand animal pathogens ( 1 )･ Until recently, picomaviruse苧wereclinical specimens (nasopharyngeal swabs: 1 ,803, cerebrospi‑

classified into five genera, but two former enterovlruSeS

nalfluids: 201, stools: 179, urine: 38, Others: 36) were collected(echovims 22 and 23) have now been placed in a new genus,

from 2,105 mostly pediatric patients (93.6% of cases) aged

Parechovirus, and renamed human parechovirus 1 and 2

0‑12 years f♭r the purpose of virologlCal surveillance in(HPEVl and 2), re苧PeCtively, because of their unique

Hiroshima Prefecture, Japan. From the specimens, a total ofmolecular and biologlCal properties (2,3). Previous studies

709 Vimses were isolated in cell cultures and/or detected byhave shown that in most cases HPEVI causes infections ofviral antlgen‑specific EIA, electron microscopy, Or viral

the gastrointestinal or respiratory tract, and in rare cases cangen?me‑Specific RT‑PCR (Table 1)･ ln thevirusIPOSitive

cause encephalitis (1). Although earlier studies have shownthat HPEVl is a common human pathogen and that infection

kom 14 patients. All the patients were infants aged under 2lwith this virus occurs early in life (I,4,5), the isolations ofyears‑old and showed upper or lower respiratory illness withthis virus were less frequent than those of other enteroviruses

fever, and diarrhea was noted in two patients (Cases No.4and 13) (Table 2). Interestingly, all of the HPEVl isolates
were isolated byusing BGM (buffalo green monkey kidney)speclmensShovm in Table 1 , HPEV 1 was isolated sporadically(6). In this paper, we report 14 cases of acute respiratory*Corresponding author: Fax: +8 1 ‑821252‑8642, E‑mail: takao@cells as shownin Table 2. In a previousstudy, Birenbaum etal. reported that BGM cells are susceptible for the growth ofurban.neJP

36

Jpn. J. Infect. Dis., 54, 2001
Table 1 ･ Summary of clinicalsymptoms and isolated/detected viruses舟om the patients in virologlCal

surveillance &om September 1 999 to December 2000 in Hiroshima
Clinical symptoms Number of Viruses isolated I ) Or detected2)
andsyndromespecimens(Numberofpositivespecimens)

Infl uenza

378

Upper resplratOry illness

715

INFLU(199), ADENO(4), MUMPS(1), ROTA(1)ADENO(I 17), ENTERO(64), rNFLU(21), HPEV(9),Lower resplratOry il lness

404

ENTERO(14), ADENO(1 1), lNFLU(17), HPEV(6),PharymgoconJuCtival fever

33

ADENO( 1 2), ENTERO(3)HSV(9), MUMPS(1), ROTA(I), SRSV(1)

HSV(1), MEASLES(1), SRSV(1)Hand‑fわot‑arld‑mouth disease

3 8

Herpanglna

22

ENTERO(5 )ENTERO(30)Exanthems

64

MEASLES(6), ENTERO(19), rNFLU(2), SRSV(I)

Mumps

29

MUMPS(1 5), ENTERO(1)AseptlC menlngltlS

1 64

Encephalitis

59

INFLU(6), ADENO(1), ENTERO(1), HSV(I), Ron(1)Gastroenteri ti s153

SRSV(3 1), ROTA(12), ADENO(7), ENTERO(4)Otb ers

I 96

ENTERO(36), MUMPS( I 9), ADENO(3)ENTERO(9), HSV(8), INFLU(6), ADENO(5),SRSV(2), MEASLES(1), MUMPS(1)I)Several ceIHines (BGM, HEp2, RD‑1 8S, FL, Vero and MDCK) were used forvirus isolation.
2) Ⅵrus antlgenS Were detected bythe specific EIA or electronmicroscopy,andvirus genomes were detectedby the specific RT‑PCR.
ENTERO: enteroviruS, HPEV: human parecoviruS, rNFLU: innuenza virus, ADENO: adenoviruS,

HSV: herpes simplexviruS, MUMPS: mumpsviruS, MEASLES: measlesviTuS, SRSV: small round shlCturedVlruS.

Table 2. Characteristics of the patientswith isolated HPEVl kom September 1 999 to December 2000 in Hiroshima
caseNo･I

Sex

male

(yeu譜C.nth)

ly,

2male

3malc

4Oy,

male
male

6female

male

8male

I

01I

I

fever (39.OoC), upper respiratory illness

BGMfcver, bronchitis

BGM, HEp2

throatswab

May 28, 2000

fever(38.OoC), bronchitis

BGMstooll)

June 22, 2000

fevcr, upper respiratory illness, diarrhea

BGM

July 2, 2000

fever (40.OoC), upper respiratory illness
fever (39.7oC), upper respiratory illness

y,5m

nasal

swab

July I 4, 2000

nasa一swab

2mm

swab

Aug. 2, 2000

fever, upper resplratOry illness

BGM, Veto仙roatswabAug. 4, 2000

fever (39.OoC), upper respiratory illness

BGMBGM1m

血roatswab

Aug. 22, 2000

ly,2mthroat

swab

Oct. 29, 2000

fever (40.OoC), bronchitisBGMthroatswab

Nov. 10, 2000

fever (40.OoC), bronchitisBGMthroatswab

Nov. 16, 2000

fever (40.OoC), upper respiratory illness

BGMthroatswab

Dec. 9, 2000

y,2male13female

Om

Oy,

7m

ly,2m

stoo121

4BGM
BGM, Ⅵ汀O

1

1

1

S:1.1aBndea諾thvl

Sept. 8, 1 999

throatI

sympb‑sOct. 20, 1 999

3m

O≠

Oy,

1m

Clinical

swab

swab

2y,2y,male
male

II

male

910ms｡ccimDeantec.0.tee.ion

血roat

nasal

10mOy,

57OmOy,speci‑ensfemale

Iy,2m

nasal

swab

Dec.

Dec.

I

9,9,

2000

fcvcr (38.OoC), upper respiratory illness, bronchitis

fever (38.OoC), bronchitis, vomiting, diarrhea

2000

fever,

upper

respiratoryillness

BGMBGM

BGM

HCoxsackievirus AIO was also isolated fromthe same stool specimen with RD‑18S cells.
21Norwalk‑like virus was also detectedfromthe same stool specimen by RT‑PCR and electron microsopy･

HPEVl (7), and our present findings confirmed that BGM

68, 8232‑8238.

cells were suitable f♭r HPEVl isolation. ln addition, BGM

4. Sato, N., Sato, H., Kawana, R.andMatumoto, M. (1972):

cells are sensitive for coxsackiev.iruses (8･9)I Thus, BGM cells
might be a useful cell line for vlrOloglCal surveillance.

Ecological behavior of 6 Coxsackie B and 29 Echo
serotypes as revealed by serologlC Survey Of general

population in Aomori, Japan. Jpn. J. Med. Sci. Bio1., 25,355‑368.

REFERENCES

5. Joki‑Korpela, P. and Hyypi岳, T. (1998): Diagnosis and1. Stanway, G. (1990): Structure, function and evolutionepidemiology of echovirus 22 infections. Clin. Infect.

Dis., 26, 129‑136.
6. Editorial Committee of lnfectiousAgents Surveillance

ofpicomavimses. ∫. Gen. Ⅵrol, 71, 2483‑2501.

2. Stanway, G. and Hyypia, T. (1999): Parechoviruses. J.

Report ( 1 997): Isolation/detection of virus from humanⅥro1., 73, 5249‑5254.

3. Stanway, G., Kalkkinen, N., Roivainen, F., Ghazi, F., Khan,sources, by year, Japan, 1991‑1996. Jpn. ∫. Med. S°i.

M., Smyth, M., Meurman, 0. and Hyypia, T. (1994):

Bio1., 50 (suppl.), Slll‑Sl 12.

Molecular and biologlCal characteristics of echovims 22,

7. Birenbaum, E., Handsher, 氏., Kuint, ∫., Dagan, 良.,a representative of a new pICOmaVirus group. J. Virol.,Raichman, B., Mendelson, E. and Linder, N. (1997):

37

Fig. 1. PFGE pattems of BlnI‑digests of chromosomal DNA of Salmonella Enteritidis isolates from 28 cases of food poisonlng Outbreaks.
(A) Salmonella Enteritidis from food poisoning outbreaks (cases 22‑28) in Hyogo Prefecture in 2000.

Lanes 1,7, and13: A DNA ladder. Lane 3: Hyogo‑SE160 (genotype A‑a) from a patient in case 22. Lane 4: HyogoISE172 (A‑a) from thecausative food, omelet, inthe same case. Lane 5: HyogoISE177 (nondescript) in case 23. Lane 6: Hyogo‑SE178 (nondescript) in case 24. Lane
8 : Hyogo‑SEI79 (A‑a) in case 25. Lane 9: Hyogo‑SE187 (A‑b) in case 26. Lane 10: HyogoISE190 (A‑a) in case 27. Lane I 1 : Hyogo‑SE193(A‑a) in case 28. Lanes 2 and 12: Hyogo‑SE186 (A‑b) and HyogoISE189 (A‑b): two isolates from sporadic cases 53 and 54 in 2000 as ref. 7.
Chr: chromosome.

(B) Representative PFGE pattems of Salmonella Enteritidis isolates in food poisoning outbreaks in llyogo Prefecture in I 997‑2000.

Lanes 1, 8, and 15: A DNA ladder. Lane 2: Ilyogo‑SEO17 (genotypeA‑a) in case 6. Lane 3: Nishinomiyal01 (A‑b) in case 2. Lane 4: Hyogo‑
SE103 (A') in case 14. Lane 5: Hyogo‑SEOO8 (A') in case 5. Lane 6: Hyogo‑SE127 (A') in case 17. Lane 7: Hyogo‑SEO65 (B) in case 10.
Lane 9: Hyogo‑SE151 (C) in case 20. Lane IO: Kakogawa‑AO3 (D) in case 1. Lane ll: Sumot0‑21 (E) in case 3. Lane 12: Hyogo‑SE177(nondescript) in case 23. Lane I 3: Hyogo‑SE178 (nondescript) in case 24. Lane 14 : Hyogo‑SE193 (A‑a) in case 28. Chr.: chromosome.
*Co汀eSpOnding author: Fax: +8 1‑78‑53 1 ‑7080

38