Jpn. J. Infect. Dis., 53, 2000

Laboratory and Epidemiology Communications
Comparison by Pulsed‑Field Gel Electrophoresis of Salmonella

Enteritidis Genotypes from Various Food Poisoning Outbreaks
from1997 to 1999 in Hyogo Prefecture
Kokichi Hamada*, Hidetaka Tsuji, Hidemasa Izumlyal and Haruo Watanabel
Division ofMt'crobiology, Hyogo Prefectural Institute of Public Health,Arata‑cho 2‑I‑29, Hyogo‑ku, Kobe 652‑0032 andJDepartment of Bacteriology, National Institute ofInfecfious Diseases,
TToyama I‑23‑I, Shinjuku‑ku, Tob70 I 62‑8640, Japan
Communicated by Shunsuke Imai(Accepted March 24, 2000)

Salmonella serovarEnteritidis has become the most preva‑

such as 1, 4, 5, 6, 6a, 34, and RDNC (Reaction Does tiot
Conform). All were prevalent in Japan (1), but RDNC (4)lent among the Salmonellae serotypes in Japan since 1989
(1). From 1997 to 1999, Hyogo Prefecture experlenCed 21

genotypes of representative isolates from the 2 1 outbreaks

emerged abruptly in 1998 in Hyogo Prefecture･ They were
examined by pulsed‑field gel electrophoresis (PFGE) byusinga Gene Path Typing System (Nippon Bio‑Rad, Tokyo) (Fig･I, 2). There were five PFGE pattems revealed by the BlnI
chromosomal DNA digests. There was no correlation between(2, 3) (Table).

PFGE pattemand phagetype.

known food poISOnmg Outbreaks caused by the serovar. To

detemine the genetic characters of the strains involved inHyogo Prefecture, we determined the phagetypes andThe 2 1 isolated strains showed various phage types (PTs),

A major PFGE pattem, group A, exhibited one PT 1 (Hyogo‑Tablet Food poisoning outbreaks of Salmonella Enteritidis (1997‑ 1999) in Hyogo Prefecture and causative strainsused inthis paper
Causative strain used

DateOccasion

ExposedPatientsst‑inI

June1997

Lunchataprlmary

SChoo12Dec･ 1997

3Feb. 1998

Funeralmeal ataninn

4Febl 1998

Lunch and/or employees at anursery

5July 1998

6Aug･ 1998

7Aug･ 1998

80ct. 1998

Nov･ 1998

12Lunchatanursery

Nov. 1998

Dinneratahotelll Dec. 1998

Lunch fordelivery

12Jan. 1999

Hospital supper

13Mar. 1999

14July 1999

15Aug. 1999

ChinesenoodlesatarestauranI

16

Sept. 1999

Creambunsoldbytraveling

In‑hospital infection

KoreanpancakeBomamarketin‑ 4‑41‑Z9m等等〝5SD A E AガA A A A B A A Aガ

S umot0‑2 180〜96641

UnboU

Kakogawa‑AO3
Nishinomiya‑0 1wn㈹EggBom astore athome

Funeral mealathome (egg dish)0ノ 2 232Ltmchatahote1
Rolled sushi Bom asupermarket

910664Dinnerathome (egg);h,T :yFGB?nIPaaem Reference

Hyogo‑ SEOO 5

7Hyogo‑SEOO

9Hyogo‑SEO 1 7

81Hyogo‑SEO2 675

3 0Hyogo‑SEO2 8111

42

Hyogo‑SEO515CCCCC

81 1

Hyogo‑SEO65

Hyogo‑SEO72*48524757

Unknown

5Hyogo‑126Hyogo‑SE 1 03Hyogo‑SEO73
SEO96

3 families
2Unknown

2Hyogo‑SE 1 06

RDNC

A206

Hyogo‑SEl 16RDNC

A

4Hyogo‑SE 1 27

cars in Hyogoand 2 Prefectures
l70ct. 1999

0meletwithrice Bom asuper‑

45A'market at home
1819Aug. 1999

Sept. 1999

Dimerathome

SushifordeliveryBomthesame

55Hyogo‑SE 1 3 8

4 1

Hyogo‑SE 1 29

RDNC

77Hyogo‑SE1 5 1I29

8Hyogo‑SE 143

RDNC

246

4AA

sushi‑bar in 3 school field days

and a wheel‑chair marathon race
20

Sept. 1999

DinnerwithafamiIy aIanIndianC

restaurantand/or employees dlere
21Sept. 1999

Hospital supperA

書A separete expenment indicated thatthe PFGE pattemswithbothBlnl and XbaI were essentiallythe same as dlOSe Of Hyogo･SEO58 in case 9･*Corresponding author: Fax: +8 1 ‑78･53 I‑708025(3)Jpn. J. Infect. Dis., 53, 2000
(A)

Fig. I. PFGE pattem ofBlnI‑digests of chromosomalDNA of Salmonella Enteritidis isolates･(A) Salmonella Enteritidis isolates with RDNC phagetypes.
Lane 1;入DNA ladder. Lane 2; HyogoISEO17 (RDNC), case 6. Lane 3; Hyogo‑SEO26 (RDNC), case 7･ Lane 4; Hyogo‑

SEO28 (RDNC), case 8. Lane 5; Hyogo‑SEO65 (RDNC), case 10. Lane 6; Hyogo‑SEO96 (RDNC), case 13･ Lane 7; Hyogo‑
SE106 (RDNC), case15. Lane 8; Hyogo ‑SEl16 (RDNC), case16. Lane 9; Hyogo‑SE129 (RDNC), case 19. Lane 10;

Hyogo‑SE143 (RDNC), case 21. Lane ll; Hyogo‑SEOO5 (PTl), case 4. Lane 12; Sumot0‑21 (PT4), case 3･ Lane 13;
Hyogo‑SEO73 (PT4), case 12. Lane 14; Hyogo‑SE103 (PT6a), case 14. Lane 15; Kakogawa‑AO3 (PT34), case l･(B) Salmonella Enteritidis isolateswithother phagetypes.
Lane 1, 15;入DNA ladder. Lane 2; Nishinomiya‑01 (PT4), case 2. Lane 3; Hyogo‑SEOO5 (PTl), case 4. Lane 4; Hyogo‑SEOOS (PT6), case 5. Lane 5; Hyogo‑SEO58 (PT4), case 9. Lane 6; Hyogo‑SEO73 (PT4), case 12. Lane 7; Hyogo‑SE127(PT5), case17. Lane 8; Hyogo‑SE138 (PT4), case18. Lane 9; Hyogo‑SEIO3 (PT6a), case 14. Lane 10; Kakogawa‑AO3(PT34), case I. Lane ll; Sumot0‑21 (PT4), case 3. LaJle 12; Hyogo‑SE151 (PTl), case 20. Lane 13; Hyogo‑SEO17 (RDNC),
case 6. Lane 14; Hyogo‑SEO65 (RDNC), case 10.

Fig, 2. PFGE paLttem OfXbaI‑digests of chromosomal DNA ofSalmonella Enteritidis isolates.

(A) Salmonella Enteritidis isolates with RDNC phagetypes.
Lane 1;入DNA ladder. Lane 2; Hyogo‑SEO17, case 6. Lane 3; Hyogo‑SEO26, case 7. Lane 4; Hyogo‑SEO28, case 8･ Lane

5; Hyogo‑SEO65, case 10･ Lane 6; Hyogo‑SEO96, case 13･ Lane 7; Hyogo‑SEIO6, case151 Lane 8; Hyogo‑SE116, caSe16･Lane 9; Hyogo‑SE129, case 19. Lane 10; Hyogo‑SE143, case 21.

(B) Salmonetla Enteritidis isolateswith other phagetypeS.
Lane 1;入DNA ladder. Lane 2; Nishinomiyal01 (PT4), case 2. Lane 3; Hyogo‑SEOO5 (PTl), case 4. Lane 4; Hyogo‑SE151(PTl), case 20. Lane 5; Sumoto121 (PT4), case 3. Lane 6; Hyogo‑SEO58 (PT4), case 9, Lane 7; Hyogo‑SEO73 (PT4),case12. Lane 8; Hyogo ‑SE138 (PT4), case 18. Lane 9; Hyogo‑SE127 (PT5), case 17. Lane 10; Hyogo‑SEOO8 (PT6), case
5. Lane 1 1; Hyogo‑SE103 (PT6a), case 14. Lane 12; Kakogawa‑AO3 (PT34), case 1.

SEOO5, lane llinFig. 1Aandlane 3inFig. lB), five PT4S

similar to group A but having an extra band unlque tO eachstrainwas named A'. It consisted of one PT6 (HyogoISEOO8,(Nishinomiya‑01, lane 2inFig. 1B; Hyogo‑SEO58, lane 5in
Fig･ 1B; Hyogo‑SEO73, lane 13inFig. lAand lane 6inFig.

case 5, lane 4inFig. lB) withanextrabandjust below 48･5
kb, one PT6a (Hyogo‑SE103, case 14, lane 14inFig･ lAandlane 9inFig. lB)withanextraband of194 ‑ 243 kb,and onePT5 (Hyogo‑SE127, case 17, lane 7inFig･ lB) withanextra
band farbelow 48.5 kb. In groups AandA', allthe non‑RDNClB; Hyogo‑SE138, lane 8 in Fig. lB;and Hyog0‑072, datanot shown), and eight RDNCs (Hyogo‑SEO17, lane 2inFig.
lAand lane 13inFig. lB; Hyogo‑SEO26, lane 3inFig. lA;
Hyogo‑SEO28, lane 4 in Fig. lA; Hyogo‑SEO96, lane 6 inFig･ 1A; Hyogo‑SEIO6, lane 7 in Fig. lA; Hyogo‑SE116,lane 8 in Fig. lA; Hyogo‑SE129, lane 9 in Fig. lA; andHyogo‑SE143, lane lOinFig. lA). The PFGE pattem quite

strains except Hyogo‑SEIO3 (case 14) had a common extra
band slightly larger than630 kb.
The remaining four stmins exhibitedunique PFGE pattems･26

Jpn. J. Infect. Dis., 53, 2000
They were classified as groups B‑E; a strainofRDNC, Hyogo‑SEO65 (case 10) as group B (Fig. lA, lane 5 and Fig. lB,lane 14); a strainofPTl, Hyogo‑SE151 (case 20) as group C

ControI Division, Minist町OfHealthand Welfare ( 1 995):

Salmonella, Japan, 1992‑ 1994. Infect. Agents Surveil‑lance Rep. 16, 1'‑2'.
2. Hamada, K., Tsuji, H., Shimada, K. and Hosoda, Y.

(Fig. lB, lane 12); a strainofPT34, Kakogawa‑AO3 (case 1)

as group D (Fig. lB, lane 10);and a strainofPT4, Sumot0‑( I 999): Epidemiologicalanalysis of Salm?nella serovarEnteritidis isolates from two food poISOnlng Outbreaks

21 (case 3) as group E (Fig. lA, lane 12andFig. lB, lane1 1). The PFGE pattems ofXbal digests were similar for all

the isolates (Fig･ 2)･ The PFGE pattem grouped as above forinthe Hyogo Prefecture. Bull. Hyogo Prefect. hst. Public
Health, 34,impress (in Japanese).

each isolate is shown in Table together withother descrlP‑

3. Tsuji, H., Shimada, K., Hamada, K. and Nakajima, H･tlOnS.

(2000): Outbreak of Salmonella Enteritidis caused by

We have thus described a PFGE analysis of Salmonella
Enteritidisfromthe recent 2 1 food poISOnmg Outbreaks. Fivecontaminated bunSpeddled by a producerusing traveling
carSinHyogoand neighboring prefectures in 1999:an

distinct pattems were detected upon PFGE. No correlation

epidemiological study uslng Pulsed‑field gel electro‑with phagetype was detected.

phoresis. Jpn. J. Infect. Dis., 53, 23‑24･

4. Ward, L. R., de Sa, J. D. H.andRowe, B. (1987):Aphage‑typing scheme for Salmonella enteritidis･ Epidemiol･REFERENCES

Infect., 99, 291‑294.

I. National Institute of Healthand Infectious Diseases

27