Jpn. J. Infect. Dis., 54, 2001

Laboratory and Epidemiology Communications

Isolation of Echovirus Type 1 3 from Patients of Aseptic MenlngltlS
Masaaki Keino*, Masahiko Kanno, Kyoko Hirasawa, Tomoko Watari, Masahide Mikawa,
Kimio Saito, Kazuo Kato, Masahiko Katayose】 and Hiromu Yoshida2

Department of Microbiology, Fukushima Prefectural Institute of Public Health,
Mitouchi 1 6‑6, Hokida, Fukushima 960‑8560,
JDepartment of Pediatrics, Public Soma General Hospital,
Tsubogasako 142, Niinuma, Soma 976‑0011 and2Department of Virology II, National Institute of lnfectious Diseases,Gakuen 4‑7‑1, Musashimurayama, To砂0 208‑0011

Communicated by Kazuo Kato

(Accepted January 3 1 , 2002)

from menlngltlS Patients among all the E13 positive cases

A district of Fukushima Prefecture experienced an outbreakor echovirus type13 (E13) in September, October, andNovember 2001. The virus was isolated from 36 patients withhad beeヮobserved previously ( 1 ,3)･

clinical manifestations such as fever, headache, resplratOry

illness, vomltlng, abdominal palm, and rash. The number ofRDl 1 8S, HEpl2, Vero, and HMV‑II cells. Ofthese cells, only
RD‑18s cells developed a clear cytopathic effect, such ascases was 8 in September, 15 in October, and 13 in November･rounding of the cells at 2 to 5血ys丘)llowed by cell lysis. TheFor vlruS isolation, the specimens were inoculated onto

cytopathic effect in Vero cells was equlVOCal･ The isolated
virus could be propagated efficiently in RD‑18s cells, andSuch a fわcal E13 Outbreak had not been reported befわre inJapan, though an isolated case was registered in 1980･
The 36 cases were chil血en of6 months to 15 years of age

the virus titer attained 106‑1 011TCID50/0.025 ml after the three(the precise ages of three cases were not recorded, however).

passages. The isolates were not neutralized by an anti‑echovirus

The number of patients in each age group from 0 to 15 years

serum pool EP95 which contained antibodies agalnSt eChoviruS

was2,4, 1,0, 5, 1,4,4, 5, 3, 1,2,0,0,0,and 1,respectively･

3,4, 5, 6, 7,9, ll, 14, 16, 17, 18,22(humanparechovims 1),

Children under 1 2 constituted the majorityin accordance with24, 25, and 30. The isolates were neutralized byanti‑coxsackie

the reports from the United States (U.S.) (1) and Germany (2)vims A6 and by anti‑A9 sera. As a consequence, we used themodified RT‑PCR with pan‑EV primer Pairs developed by

but in discordance with the situation in the United Kingdom

Centers for Disease Control and PreventiPn (CDC) (4‑6)･ By
RT‑PCR with two prlmer Palr combinations, i.e., 187, 188,
189‑011 and 040, 012‑011, PCR products of the VPl reglOn(U.K.) (3) where two‑thirds of the isolates were from patientsover 15 years of age.

The male to female ratioamong the patients was 7 to 5 in thepresent outbreak; a similartendency was observed in the reportof the enterovimses were obtained. The sequence analysis offrom the U.S. (I). However, it has not yet been establishedthe PCR products showed about 80% CDNA sequencethat the males are more prone to E13 infection.

homology with the E1 3 prototype, Del Carmen strain, and98 to 99% homology with the U･S･ and the European E13

The patients consulted pediatricians at a sentinel hospltal

isolates obtained in the last two years. The neutralization test

particIPatlng in the National EpidemiologlCal Surveillanceof Infectious Diseases in the district and were diagnosed
tonsillitis (4 cases)*, pharyngitis (3 cases)*, pneumonia (1with an anti‑E1 3 serum confirmed that the isolates were E13.
CDC reported that among 76 E1 3‑positive cases reported
from 13 states in the U.S. in March‑June 2001, 50 cases werec警e)*, bronchitis (1 case)*, tympanitis (1 case)*, fever convu1‑

diagnosed as aseptic meningitis (1). The CDR Weekly (3)sIOn (1 case)* and viral exanthema (1 case), where diagnos享sreported that among 67 E13‑positive cases in the first 30with asterisks contained cases wi血more than one diagnostlC

weeks of year 2000 in the U.K., 38 were diagnosed as viralmenlngitis. Gemany, where E13 cases were rare, reported

as aseptic meningitis (24 cases), gastroenteritis (5 cases)*,name. In the cases of aseptlC menlngltlS, a fever of 37.0‑

39.70C persisted for 2‑4 days and a headache for 4‑6 days･

an increase ofE13 meningitis (2). The source of infection ofAll patients were admitted to the hospital but discharged
without residual effects. Generally, the menlngltlS in thethe present outbreak in Fukushima Prefecture is not known;

present outbreak was relatively mild.

established. As of ll January 2002, 26 additional casesa link to a visit to the above epidemic countries cannot be

A total of 65 E13 isolates were obtained from the 36

including two cases suspected of aseptlC menlngltlS appearedin the district. The clinical specimens collected during

patients, 28from throat swab specimens, 26 from rectal swab

specimens, and 1 lfrom cerebrosplnalfluid specimens.Among

December 2001 are now under virologlCal examination.

24 menlngltlS patients, E13 was positive in the cerebrosplnallt should be mentioned that a voluntary fax from anotherfluids of I 1 cases, but not in those of other 13 patients. The

hospital in the same district informed Fukushima Prefectural

throat and/Or rectal swab specimens of the latter group were,

hstitute orPublic Health of a sporadic occu汀enCe Ormenlnglbs

however, positive for the virus. Similar isolation rates ofE1 3

already in the middle of August. The Institute took immediate
public health measures by waning district physicians or the*Corresponding author: Tel: +8 1 ‑24‑546‑7 1 05, Fax: +8 1 ‑24‑546‑

possibility of an increase in the number of meningitis patients
through the weekly report on infectious diseases･

8364, E‑mail:fukushimaeiken@mbp.sphere.ne.jp249Jpn. J. Infect. Dis., 54, 2001
4. Oberste, M. S., Maher, K., Kilpatrick, D. 良., Flemister,This article appeared in Infectious Agents SurveillanceReport (lASR), vol. 22, m0. 12, 317‑318, 2001 in Japanese.

M. 良. and Pallansch, M. A. (1999): Molecular evolutionof the human enterovimses: co汀elation of serotype withVP I sequence and application to pICOmaVirusclassifi‑cation. ∫. Ⅵro1., 73, 1941‑1948.

REFERENCES

5. Oberste, M. S., Maher, K., Kilpatrick, D. 良., Flemister,

M･ R., Brown, B. A. and Pallansch, M. A. (1999): Typing

1. Centers for Disease Controland Prevention (2001 ): Echo‑virus type 13‑‑‑United States, 2001. Morbid. Mortal.
Wkly. Rep., 50, 777‑780.

orhuman enteroviruses by partial sequenclng OfVPl. ∫.

Clin. Microbio1., 37, 1288‑1293.

2. Robert Koch Institute (2000): Eine H畠ufung viraler

6. Oberste, M. S., Maher, K., Flemister, M. 良., Marchetti,

Meningitis durch ECHO‑Virus Typ 1 3 in Sachsen‑Anhalt.

D. R., Kilpatrick, D. R. and Pallansch, M. A. (2000):

Epidemiologisches Bulletin, 38, 305‑306.

Comparison of classic and molecular approaches fわr theidentification of untypeable enteroviruses. J. Clin.

3 ･ PHLS Coヮmunicable Disease Surveillance Centre (2000):

Microbio1., 38, 1170‑1 174.

Ⅵral menlngltlS associated with increase in echovirus

type 13･ CDRWkly., 10, p. 277, p. 280.

250