9
ῌ῎ ῐ῍ Helicobacter 臆臰腇腚臐臵腢腗腨腢膪臰臥膈膄腦腊膂膌膏腴臷臤腂膧臤腂臁膖臲腂膝腶膰膖腂腅腴膥腽腂膦臖臺 膭膘膐膋腍膑膭臙臝 膓腋 20 4 21 膓腋 20 8 1 Helicobacter 臆臰腱臻腞腔腗膄腦腊膂膌膢腣腒腯腦腝臯腡 Helicobacter pylori 腡腫腭 臘腰腯腜腏腗腎腁 臇Helicobacter cinaedi 腢臐臵臛腎臢臒腓腯腮腒腞腍腬腁 H. cinaedi 膹腧腝腉Helicobacter 臆臰腱臣腉腁 膪臰臥膈膄腦腊膂膌膢腱臘腙腗腀 Helicobacter 15 臆臰膺臶膯腞 H. cinaedi 膒膮14 膯腱腉腜腁 腾腈腎膡臎膚膰腲膄腦腁 膇膊腷膂腶 腺腄膰腲膄腦膔膘腸腵膌腄膄腦膔膘腸腵膌腄 EX 膄腦腁 膙臑膠膭腢 HP 膒膮膄腦Vi 膇膊腷 腲膄腦腵腎膡腢膅膌膊膰腲膄腦腌腫腤膈腶腽膍腼腳腻腵膍腹膍腢腷膌膍膄腲 HP 膄腦8 臰腱膬腔腗腀 8 臰腢膄腦腢腊腘腁 膇膊腷膂腶腺腄膰腲膄腦Vi 膇膊腷膰腲膄腦腁 膅膌 膊膰腲膄腦腣腁 臎膣膄腦腝腈腭腁 腓腬腡膷臓臋腃腖膣腓腯腜腉腮腗腨腁 膆腷膝膄腦腢腷膌腾腄腱臉腔腁 臎膚膰腲膄腦腫腭腩臨腡腫腭腇腚腡臆腢膕腎膫腝腏腗腀 腒腢 3 臰腢膄腦腝腣膇膊腷膂腶腺腄膰腲膄腦腝腢膕腎膖膉膲臖腝腈腙腗腀 膕臆腢膮腕腮膠腱腝臘腙腗 ATP 腊腙腰腝腣腁 臎膚膰腲膄腦腌腫腤膔膘腸腵膌腄膄腦臼腡膕腔腗 腢膮腊腙腰腤膄膩 5 腾膠腡膾臌腡膟腕腮腢腡腔腁 膇膊腷膂腶腺腄膰腲膄腦臼腢臆ATP 腣膓自腓腯腜腉腮腒腞腎腬腍腡腠腙腗腀 腓腬腡臞腢臉膶臠腱臘腊腞腁 臎膚 腲膄腦腌腫腤膔膘腸腵膌腄膄腦臼腢臆腐腎膄膩 5 腾膠腡腣臀臆臉腡膠腔腜腉腮腢腡 腔腁 膇膊腷膂腶腺腄膰腲膄腦臼腢臆膪腓自腓腯腜腉腗腀 膏臼腢臍膥腍腬腁 膇膊腷 膂腶腺腄膰腲膄腦腇腚臐臵腁 膕臦自膱膟腡腌腉腜腯腜腉腮腞臗腋腬腯腗腀 Key words: Helicobacter cinaedi腔腣膄腦腇腚臐臵 Helicobacter 2008 4 臏臔臟 30 臆臰腡膒膵 腓腯腜腌腭腁 腖腯腬腣 腏腐 膵臟膐臆 (entero- hepatic) 腞膔臟膐臆 (gastric) 腑腬腯腮腀 膃腽腍 膒膮腓腯腮 Helicobacter species 腞腔腜腣腁 gastric Helicobacter species 腡膹腦腯腮 Helicobacter pylori 膣膜腝腈腮腎腁 enterohepatic Helicobacter species 膒膵腓腯腮 Helicobacter cinaedi, Helicobacter bilis, Helicobacter canadensis, Helicobacter canis, Helico- bacter fennelliae, Helicobacter pullorum 腬腯腜 腉腮 1) 腀 腠腍腝腩 H. cinaedi 1980 膁腡膏臚腁 膡腬腍 腢膺臬膱腡腫腭膛膻膟腔腗膱臮腢膴腪膵 腁 臎膚腍腬腢膒膮膶臛腓腯腜腉腮 2, 3) 腀臇腣腁 膛膒腢腠腉膱臮腍腬腢臹膶膖臛腩臒腬腯腮 4) 腦腗腁 腹腰腶膌腄膍腢 H. cinaedi 膖腢膜腔腁 膘腱膗腏膽腒腔腗臛腩腈腮 5, 6) 腀 膃腽膏 膩腡腣腴膀腁 膁膉腸腺腄腠腟腢膤腍腬腩膒膮膖臛腎 腈腮腆腨臹腝腈腮腞臫腡腁 臄膲臹腞腔 腜膐腛腑腬腯腜腉腮腀 腒腢腫腊腠臊腍腬腁 H. cinaedi 臸腢臔腡腌腉腜膑腕腥腏臆臰腝腈腮 腎腁 臆臰腱膹腧 Helicobacter 臆臰腣膄膩腎臜腩腢腎腉腀 臆腢膕腣臖膼膳臃腝腉腾膸腱腔腁 腦腗腁 膤腗腊腱臉腕腮腩腢腎腐腁 膓膇腡腣 膆腳膋膉(thin spreading layer) 膕腕腮腗腨膕腢膫腎臜腝腈腮腀 腖腢腗腨腁 臸臔腝腣 Heli- cobacter 臆臰腢腇腚腍腚膫臭腠膄膩膗膘腢膫腎膿 腨腬腯腜腉腮腀 Helicobacter 臆臰腱臻腞腔腗腔腣膒膮膄腦腣腁 臔臟臰腎臥腓腯腜腉腮腎腁 腖腯腬膄腦腊膂膌膷膫腐(4648650) 臕膞臥臰臈1100 膭膘膐膋腍膑膭臙臝 臖臺 TEL: 0527576780 FAX: 0527576780 E-mail: [email protected] 腾膚膴膋腍膑膭臡臩 Vol. 18 No. 4 2008. 腾膚膴膋腍膑膭膨 2008 7 227

Helicobacter - 一般社団法人日本臨床微生物学会 xx xx xx w xx ww w H. canadensis 600T ANE xx x wwxyyy CAM x ww wxxww H. canis 598T ANE xx xx x x xx y y y CAM xx x x x

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Page 1: Helicobacter - 一般社団法人日本臨床微生物学会 xx xx xx w xx ww w H. canadensis 600T ANE xx x wwxyyy CAM x ww wxxww H. canis 598T ANE xx xx x x xx y y y CAM xx x x x

�� ��

Helicobacter�������������������

��� ����� � �� ���� ���� �������������������

��� 20� 4 � 21���� �� 20� 8 � 1��

Helicobacter���!!"#$������%&'()*+ Helicobacter pylori +,-./'01�2� 3�� Helicobacter cinaedi ���"42567'8&#9:� H. cinaedi !;<#$� Helicobacter���!=>� ��������!.?�@ Helicobacter� 15

��ABC# H. cinaedi%D&'C 14 C!(>0� �)*��+EFG,��� HIJKLMNG,���-.OPQN���-.OPQN EX��� RST�� HP&'��� Vi HIJG,��� U/*��VQIG,��� W,XYLZ[\]^P[_[�JQ[`a HP G,��� 8 �!0b$�@ 8 �����cd� HIJKLMNG,��� Vi HIJG,��� VQIG,��%� E1;2��)e-� 7:+fgh3425i7'0>8���67�j��+k3�JQlN!m�$� EFG,��,-n8o+,-��+��9p2q:)1�@ &� 3

�����;)%HIJKLMNG,��)�9p2�<=�)e?�@ 9p��r�!s88>).?� ATP r�?@)%� +EFG,��W,X-.OPQN��t+9p$��A�r�?@B2�C 5�8+uv+Dws8�+!$� HIJKLMNG,��t��A� ATP r�%xy7'0>8&#2�:9+j?�@ 7:+zE�mF{|!.c#� +EFG,��W,X-.OPQN��t��A�#}2�C 5�8+%~�m+-T$0>8�+!$� HIJKLMNG,��t��A%GHmF2Iy7'0>�@ �t���9:� HIJKLMNG,��%����� 9p�yJ�KL+W>0M'0>8#��:'�@

Key words: Helicobacter cinaedi� NO��� ����

� �Helicobacter �% 2008� 4 ��� 30 ��+&P

7'0W-� �':%�1}Q���� (entero-

hepatic) #���� (gastric) +&�:'8@ �Z9:&'7'8 Helicobacter species #$0%� gastric

Helicobacter species +;('8 Helicobacter pylori

22R)e82� enterohepatic Helicobacter species

+&P7'8 Helicobacter cinaedi, Helicobacter bilis,

Helicobacter canadensis, Helicobacter canis, Helico-

bacter fennelliae, Helicobacter pullorum n�:'0>81)@ j9)n H. cinaedi % 1980�S��� �:9

�AT��+,-U���2Dw$����Q���V� EF9:�&'W2"47'0>82, 3)@ 3�)%� U��X�j>��9:��W"4n6:'84)@(�� Y@LQN[� H. cinaedi 2Z��[�\)]�$� �\�^!�1�&$�"4ne85, 6)@ �Z��+%��� � OMNj¡�¢_9:n&'"42e8`_£a�^�)e8#Z¤+� b¥�^�#$0�c¦�:'0>8@ &�,cjd§9:� H.

cinaedi %%D��e+W>0f¨s©1��)e82� g��!;< Helicobacter���%�C2ªhjn�2#>@ ��9p%��«¬­)$�®!i$�(�� jk�¯l!m�s8n�2#}� �m+%°]± n (thin spreading layer) +�ps8��9p�q:2ªh)e8@ �����%D�e)% Heli-

cobacter������9²q³j�Co.�qp2´�:'0>8@

Helicobacter���!!"#$�NO&'��%���$�2�7'0>82� �':�����

q�rstµ (¶464�8650) R·¸u�¹vgw 1�100�������������������TEL: 052�757�6780FAX: 052�757�6780E-mail: [email protected]

�g%D����º» Vol. 18 No. 4 2008.

��g%D����¼ 2008

7

227

Page 2: Helicobacter - 一般社団法人日本臨床微生物学会 xx xx xx w xx ww w H. canadensis 600T ANE xx x wwxyyy CAM x ww wxxww H. canis 598T ANE xx xx x x xx y y y CAM xx x x x

��� H. pylori �������7)� � � H.

pylori����� H. cinaedi�������������� �!� H. pylori"#�$%���&��'���&(�) � Helicobacter* 15$%+,-.�/ H. cinaedi�0�- 14-���1�2 8%�&�(�3�' ���� 4�56(��7��

�����1. ��� 89:;�2 <�=>�? .�/ Campylobacter

spp.���2 1��@���ABCD�2 <�=>�?� ��ABCD�2 EX <�=>�?�H. pylori����2�E�FGHIJKD:;�2 <�=>�?� LMNOPM HP��2 <QRST?� LMNOPM ViFGH:;�2 <QRST?� HCUVM HP:;�2 <�� BD?� WCG:;�2<XY>�? � 8%��2(�3 �� 89:;�2� ��ABCD�2� ��ABCD

�2 EX� LMNOPM HP��2� .�/HCUVM HP:;�2� 5%��2Z��89([� .�� Helicobacter*$%�\]�2^��_�]�`�(aZ7�� b�� FGHIJKD:;�2�LMNOPM ViFGH:;�2� WCG:;�2�3%�cd8e([� ��_��2�E�� �]f� tetrazolium violetghi��@��j$k��l��@m��n=opo���qr��� s](t7��

2. ���Helicobacter * 15 $ % + , -u H. bilis PAGU

599T (vLMG 18386T), H. canadensis PAGU 600T

(vCCUG 47163T), H. canis PAGU 598T (vNCTC

12379T), H. cinaedi PAGU 597T (vCCUG 18818T),

H. fennelliae PAGU 601T (vCCUG 7546T), H. pullo-

rum PAGU 602T (vNCTC 12824T), Helicobacter an-

seris PAGU 940T (vMIT 04�9362T), Helicobacter

brantae PAGU 941T (vMIT 04�9366T), Helicobacter

marmotae PAGU 943T (vMIT 98�6070T), Helicobac-

ter mastomyrinus PAGU 944T (vMIT 97�5574T),

Helicobacter ganmani PAGU 603T (vCCUG

43526T), Helicobacter hepaticus PAGU 604T (vLMG

16316T), Helicobacter muridarum PAGU 606T

(vLMG 13646T), Helicobacter pametensis PAGU

607T (vLMG 12678T), Helicobacter acinonychis

PAGU 609T (vLMG 12684T)(�@���!�� H. cinaedi�w�.�/w#��0�-

14-�x� ��

3. � �������a) 8� ��������������������yAz{|}UID��� �M~C��J��� �B}UW�c|��� 3%(�@�� �yAz{|}UID���� ���(n��� N2 80��CO2 10�� H2 10����yA�z{ � (Fig. 1)� �!���P����D�D(|}UID��z@.� ��(�� �� �M~C��J���� M~C��JFGH���&$������ <� yAST? ���}D <� yAST? (x� � � !"���(�� �� �� �B}UW�c|���� BBLB}UW�c|����A�� (BD)�#*�9k(�$ � �c|(%& �� �� �'�g!����B}UW�c|��(g����@��yAz{|}UID���(�@� 8%�1�2� Helicobacter* 15$%+,-.�/ H. cinaedi

�0�- 14-( 37�� 3� �� � $��l¡¢()£¤¥��� 3¦§��' �� ¨�� M~C��J���� B}UW�c|����.@�� ©ª�!�����E� enterohepatic Helicobacter

6$%+,-(�@� !"��' ��

Fig. 1. Gas displacement chamber system.�chamber (desiccator with valve), �pressure gauge, �vacuum pump, �compressed gas cylinder (N2 80�«CO2

10�«H2 10�)

*¬­®�¯®¬°��±²b³�´µ+b�¶¬,-�·¸�¹*¬­®�¯®¬°��±²b³�´µ+b�¶¬,-�·¸�¹

���0�º.T»¼ Vol. 18 No. 4 2008.8

228

Page 3: Helicobacter - 一般社団法人日本臨床微生物学会 xx xx xx w xx ww w H. canadensis 600T ANE xx x wwxyyy CAM x ww wxxww H. canis 598T ANE xx xx x x xx y y y CAM xx x x x

b) ���������� ������� ���� ������� � ���� ��������������� �������� !�"# $%� &���'()*+ �,-# ./012�3��456789�� �������:;<=>?@AB9���� ������� � 3

C����DE"�� HelicobacterFGCHIJKLMNOP�QRS�T# �UVMWX�YXZ[�\S;] 200 ml�^GW�_`a��Mbc# ��������d�U"� H. cinaedi PAGU 597T�_`a��Mbc�^GW�ef"# g�hhijklmn�� o6pqrst@9� 37u# 3vw�U"# 1

vx�MIJyz�{|"��

c) ATP�����������������$%�IJ`a�IJ012�3��# �������# 456789��# =>?@AB9�����3C����DE"�� d�U"� H. cinaedi PAGU

597T:;<}~���X���3S PAGU 934�D�# ��9�9�t�t�86 (MHB)M� McF#1

�G����"# ��Mg�G�� 10���"��_��M��"�G�� 200 ml���C"�� �9������Mklmn# o6pqrst@9�E�� 37u# �2�# O�y���U"�� �U 1v�#3v�# 5v�M_��M�J"�G�� ATP '�¡¢"�� ATP '£H# _��MMHB 3 ml�bc��9��G��¤¥�c# G��R¦�§¨"

Table 1. Growth level of each media using gas displacement chamber.

SpeciesPAGU

No.

Blood Non blood

�Bloodagar

�Skirrow �SkirrowEx

�HPselective

�ColumbiaHP

�Helico-bacter

�ViHelico

�Pyloriagar

H. bilis 599T ©© ©© ©© ©© ©© ©© w ©©H. canadensis 600T ©© ©© ©© © ©© ©© w ©H. canis 598T ©© ©© ©© ©© ©© ©© w ©H. cinaedi 597T ©© ©© ©© ©© ©© ©© © ©©H. fennelliae 601T ©© ©© ©© ©© ©© ©© © ©©H. pullorum 602T ©© ©© ©© ©© ©© © w ©

H. anseris 940T ©© ©© ©© w © ©© © ©©H. brantae 941T ©© ©© ©© ©© ©© ©© w ªH. marmotae 943T ©© ©© ©© ©© ©© ©© w ©©H. mastomyrinus 944T ©© ©© ©© ª © ©© © ªH. ganmani 603T ©© w w © w w w wH. hepaticus 604T © ©© ©© © ©© w ©© ©H. muridarum 606T ©© © ©© w w ª ª ªH. pametensis 607T ©© ©© ©© ©© ©© ©© w ªH. acinonychis 609T ©© ©© ©© ©© ©© w © ©©

H. cinaedi 611 © ©© © ©© © ©© w ©©H. cinaedi 612 ©© ©© ©© ©© © © ª wH. cinaedi 615 ©© ©© ©© ©© ©© ©© © ©H. cinaedi 616 ©© ©© ©© ©© ©© ©© w wH. cinaedi 623 ©© ©© ©© ©© ©© ©© © wH. cinaedi 624 ©© w w w w ª ª ªH. cinaedi 630 ©© ©© ©© ©© ©© ©© © ©H. cinaedi 636 ©© ©© w ©© © © © ©H. cinaedi 640 ©© ©© © ©© © © w wH. cinaedi 641 © w w w w ©© w ©H. cinaedi 642 ©© ©© © ©© ©© ©© © ©©H. cinaedi 824 ©© ©© ©© ©© ©© ©© © wH. cinaedi 825 ©© ©© ©© ©© ©© ©© © ªH. cinaedi 934 ©© © © © ©© © w w

Growth level was evaluated in four stages. ©©: good growth, ©: middle growth, w: weak growth, and ª:no growth.

HelicobacterFGC«¬���)*HelicobacterFGC«¬���)*

v­����®¯°± Vol. 18 No. 4 2008. 9

229

Page 4: Helicobacter - 一般社団法人日本臨床微生物学会 xx xx xx w xx ww w H. canadensis 600T ANE xx x wwxyyy CAM x ww wxxww H. canis 598T ANE xx xx x x xx y y y CAM xx x x x

��� �� 50 ml���� Bac Titer-Glo Microbial

Cell Viability Assay (Promega) 50 ml�� �������������� (DTX 880. Backman

Coulter)������ �!"��#$%&'( 1)*� 3)*� 5)*&+,-&./

0'1���234�5678�9:;<=&.�>?�#

� �1. 8�������� �������3@�'(AB� 3)C'(���� D2��3E

�FG��# 8@�HI'1&./JKLM�NOP�'(��QR� Table 1, Fig. 2&ST# U�� VWXYZ�[.\]NO^Y_�'(�QR� Table

2&ST# `ab�cd�� 3@�'(e�f�gJKLM�NOP���'(hij�2kl�m?�#nopqr'1�st 7@�uv'1�f�g� we

Fig. 2. A summary of the growth level in eight media using gas displacement chamber.Number of strains evaluated by each growth level (xx, x, w, y) was showed.

Table 2. Growth level using Aneropack and Campypouch.

SpeciesPAGU

No.Gas

system

Blood Non blood

�Bloodagar

�Skirrow �SkirrowEX

�HPselective

�ColumbiaHP

�Helico-bacter

�ViHelico

�Pyloriagar

H. bilis 599T ANE x xx xx w xx w w wCAM xx xx xx w xx w w w

H. canadensis 600T ANE xx x w w x y y yCAM x w w w x x w w

H. canis 598T ANE xx xx x x xx y y yCAM x x x x x y y y

H. cinaedi 597T ANE xx xx xx xx xx xx x xCAM xx xx xx xx xx x x x

H. fennelliae 601T ANE xx xx xx xx xx xx xx xxCAM xx xx xx xx xx x x x

H. pullorum 602T ANE xx xx x x x y y yCAM x x x x x w y y

Each type strain of six species designated as an enterohepatic Helicobacter species was used. Growth levelwas evaluated in four stages, same as the legend in Table 1 (xx, x, w, y). ANE: Aneropack, CAM:Campypouch.

z{|}~�}{��~����~����~�{��~��l�z{|}~�}{��~����~����~�{��~��l�

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���������� ����� ��������� ������ � !"# �$%��&'()*+,-�./��� 012341 Vi()*./��� 5�)./��� 36�78���9�:� ()*+,-�./���� ���;�<!"#

2. ���������� �()*+,-�./��:� =��./��>?@�������ABC 1DE>�FGHCI�JK 8L�LM�N" (Fig. 3)# ()*+,-�./���:OPQRRS TUVWCI'"X� ��Y�Z� [&'>78��Y�\R�]^%_`H"# aW�bc� ()*+,-�./��:defg24efh ��i��LM jk�<!"# l"����?m�noHp&I>I�qr stW"#

3. ATP�����������������=��./��ubv?@�������H"��� ATPwxyz%{!">a|� �i 3D}~����� ��H"# ��� ()*+,-�./�������yz�:� ��� (PAGU597): 3

D}~��p ���H� ����� (PAGU934)�: 5D}l� ATPwx�:Y�H" (Fig. 4)#

ATPwxyz>�e�� ������%noH">a|� =��./��ubv?@������Y���:�i 5D}�:����_ (coccoid form)��!CI"���H� ()*+,-�./��Y���: 5Df�i����_%��HCI'�� � !" (Fig. 5)#

Fig. 3. Comparison of the color change by the colony growth.Bacterial growth was observed at daily intervals for three days. H. cinaedi PAGU 597T wasincubated in a gas displacement chamber. Purple color depending on bacterial growth was easyto find on the Helicobacter agar from the first day.

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� ���� ����������� ������

�������������������� !�!� Helicobacter�"#�$%&'���()!*++� ,!�"� CFU (Colony Forming Unit)

*-.�����/0��1�2� 3��� �45��� �67�8*9:;<���/0�=*ATP >?���!� @"�A"�>?B ���!<� ATP >? *-.� 3�����,CD�����/�� �EFG���H-I��JK���L�"���� 5��*6���M>?��N!� �+�O"(*�P!<;� Q�� RS�TU8�G���L�"�� 5�V��WX�M>?��N���+� �Y*� "�XYZ[(��D!<;�X���+\Y�� H. pylori ����]!;^_* ����YZ[(�YO"(*�P�����!Y�<;� ("��P*�`#� a#� bJcd� $e�����6f��ghg6cd�i%!<;��� O"(j�("�P�kl�!< “nonculturable

state” j�m/�2.� “bacterial cell death” �&n�2��;0'o�2�8) H. cinaedi X()�,*?�2����p+*qrY�� !���<s��/0t*� u*O"(*�P!<;�"��,"!

<X-v!<�6;wx?�2.� .y�/0�2� ��-06z1�Yqr��� RS�TU8�G����2{V��WXYZ[("�|D��<H.� 3���-.X,CD��4�<;��qrY�� ���}#~���=*s���5�/06���*H;<�7�*89�2������ g��s�{�,^_*H;<�����+ *X.y����2� Helicobacter�"#���*�a#�/0�!� ����+ ��;�"��,!6; �:����9��t� ;<=��>!<�Y��!�{V*-�<����?@#��P���=� (�����9!<X��*)�����X�A����X2� ��BC�DE*F��=� �45������t*��L*G"�� 200 ml �Z<�Y"��H#���;0���9;� ��*-.� p+6�+ ������ s�{�,�"X�I�������� �45��9!� 3 #�����J��� bJ �

��%T�*-�����X,K��2��;0kl�LY�� bJ ���%T�����M�W�N2 80 � CO2 10 � H210 �bJ� �!<;� Q��¡¢£¤UH-IK�%¥£¦�� O2 �§¨! CO2 ����$J©ª��=� ¡¢£¤U

Fig. 4. Evaluation of bacterial activity using ATP assay system.H. cinaedi PAGU 597T or PAGU 934 (clinical isolate) was cultured on each medium for 5 days.Bacterial cells were collected from each medium, on day-1, -3, and -5, and measure the activity.Data were means of triplicate. The bacterial activity of the cells collected from Blood agar andModified Skirrow agar after 5 days culture were dramatically decreased, whereas the cells fromthe Helicobacter agar were still increasing or slightly decreased. RLU: relative luminescenceunits.«¬« : Blood agar, �­

: Skirrow agar, ��®� : Helicobacter agar.

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�� O2� 6�12�� CO2� 5�8������ �������� O2� 5�15�� CO2� 5�12������ ��������������� !"� 2#�� $!�%&'�()���*+�,�� Bergey’s Manual of Systematic Bacteriology� -H. pylori, H. mustelae, H. nemestrinae, H. acino-

mychis�./012�,34()5678�9: !;<=3>9��� ?@ -H. cinaedi��� ()�ABCD,�9�AB5EFG�: !;H=3>9�9)� IJ� ()���KL� HelicobacterMN#�ABOPG�!9QRS10)T,�� UV�WX�Y3Z!?[8J� �����\]^�_�� `ab�

� c�de�f� ghi��j�klZ�����������]�md5no8J�� Y�" BDpOxoid�q�r0���st]�md5u_v�w�8�xo8� ()5yz��{|5}~G�@$�RS=3>9�11)� HelicobacterMN#�� ���� ��hQ�()��5A�G�� ]�md�xo���=3��

HelicobacterMN#��<8JhQ]�������5�~8��� �������d����5�~G��!��9� H. cinaedi���� H. bilisp H.

canadensis�qT���d��AB��Z3�!;H=3>9�12, 13)� ��� ����xo8J�wlZ

Fig. 5. The morphological change of the bacterial cells.H. cinaedi PAGU 597T was cultured on each medium for 5 days. Bacterial cells were collectedfrom each medium, on day-1, -3, and -5, and observed the morphology with Gram-stained undermicroscope (�1,000).The cells collected from the Helicobacter agar on the day-5 sustained the spiral form, howeverthe majority cells from Blood agar and Modified Skirrow agar changed the form into coccoidshape.

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������� 6������� H. fennelliae�� ����������� �� 5������������������ �! ���������"#��$� %&�'��()������*+�, -����.�/01� �! ���%2�'������3�� -�����4)�����5��*+6�7 -����8��9� 24:;������<��=�! > ?1@��ABC@D�� Helicobacter E���FG�HI@����$� ))J�'����KL�MNOP�� ViKL�Q��� RSLQ������,TU �! V � 3���� -�W�XY,� KL�MNOPQ���� -�Z��9��[@\�� �! ViKL�Q��D]^RSLQ��@�� _`abcde!fG&+g�"#� $chij% k&lm '(���'n ���! �� KL�MNOPQ��@�o�Nc)*'(_`ab'n ���"#&lj%'(���'n ��+�! "#&lj%'(���� p�qors�+,@'t -uv-. ���,01� �� /�@'n �wxy b+g�'0qors[��z{P�| kS�H|m +g}��c�6� >�12� -3~#�,������ -uv�@\�� ����+�[,������! �[�+�� HelicobacterE��� -uv���j@����4���9� ����5���!�3�F6���[�� KL�MNOPQ��k7��8m � HelicobacterE���HIFGD]^ -uv-�9:@D��. �D9� �;���,���/��,�<��!

� �1) Solnick, J. V., D. B. Schauer. 2001. Emergence of

diverse Helicobacter species in the pathogenesisof gastric and enterohepatic diseases. Clin. Mi-crobiol. Rev. 14(1): 59�97.

2) Totten, P. A., F. C. Fennell, F. C. Tenover, et al.1985. Campylobacter cinaedi (sp. nov.) and Cam-

pylobacter fennelliae (sp. nov.): two new Campy-

lobacter species associated with enteric diseasein homosexual men. J. Infect. Dis. 151: 131�139.

3) Kiehlbauch, J. A., R. V. Tauxe, C. N. Baker, et al.1994. Helicobacter cinaedi-associated bactere-mia and cellulites in immunocompromised pa-tients. Ann. Intern. Med. 121: 90�93.

4) Vandamme, P., E. Falsen, J. De Ley, et al. 1990.Identification of Campylobacter cinaedi isolatedfrom blood and feces of children and adultfemales. J. Clin. Microbiol. 28: 1016�1020.

5) Kitamura, T., Y. Kawamura, T. Akaike, et al.2007. Helicobacter cinaedi cellulites and bac-teremia in immunocompetent host after ortho-pedic surgery. J. Clin. Microbiol. 45(1): 31�38.

6) Iwashita, H., S. Fujii, Y. Kawamura, et al. 2008.Identification of the antigenic protein of Helico-

bacter cinaedi and its immunogenicity in hu-mans with H. cinaedi infections. Clin. VaccineImmunol. 15(3): 513�521.

7) �� �� �=��� �� >� �� 2000. Heli-

cobacter pylori HIFG��5� �KL�MNOP��� k7��8m� �?�� �A@}#HI������ 11(1): 27�32.

8) Kusters, J. G., M. M. Gerrits, C. M. Vanden-broucke-Grauls, et al. 1997. Coccoid form ofHelicobacter pylori are the morphologic mani-festation of cell death. Infect. Immun. 65(9):3672�3679.

9) Garrity, G. M., J. A. Bell, T. Liburn. 2005. Fam-ily II. Helicobacteraceae fam. nov. In Brenner, D.J., Krieg, N. R., Staley, J. T., and Garrity, G. M.(eds.), Bergey’s Manual of Systematic Bacteriol-

ogy. 2nd ed. Vol. 2. New York, Springer-Verlag.10) A���� �BC d� �¡ �� �� 2007� %

&�[�� Helicobacter cinaediD]^>�D¢�����,£EFGH@¤��F6� ¥¦§¨©� 81: 700�706.

11) Iª«� ¬=J­� ®¯°K� �� 2005. H.

cinaedi, H. fenneliae, H. pullorum�/�,L±/� �A,@}# 32: 175�180.

12) Fox, J. G., L. L. Yan, E. N. Mendes, et al. 1995.Helicobacter bilis sp. nov., a novel Helicobacter

species isolated from bile, livers, and intestinesof aged, inbred mice. J. Clin. Microbiol. 33(2):445�454.

13) Fox, J. G., C. C. Chien, F. G. Rodgers, et al. 2000.Helicobacter canadensis sp. nov. isolated fromhumans with diarrhea as an example of anemerging pathogen. J. Clin. Micobiol. 38(7):2546�2549.

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Evaluation of Various Media for Rapid Detection of Helicobacter spp.

Junko Tomida, Mitsuaki Kashida, Kazuki Oinishi, Ryuichi Endo, Yuji Morita, Yoshiaki Kawamura

Department of Microbiology, Aichi-Gakuin University, School of Pharmacy

The evaluation of commercially available media for Helicobacter species has been carried out mainly byHelicobacter pylori strains. However the reports of Helicobacter cinaedi infection have been increasing, wedecided to investigate the performance of various media for growing several Helicobacter species. Weevaluated following 8 media with using the type strain of 15 species of the genus Helicobacter, and 14clinical isolates of H. cinaedi; Sheep blood agar (Nissui), Modified Skirrow agar (Nissui), Modified Skirrowagar EX (Nissui), HP selective agar (Eiken), Columbia Helicobacter pylori agar (BD), Helicobacter agar(Nissui), Vi Helico agar (Eiken), and Pylori agar (Kyokuto). Because the latter three media were containinghorse-serum and indicator-dye (tetrazolium salt), the bacterial growth was recognizable at a glance by theviolet-blue color on the translucent media. Within these three media, Helicobacter spp. was the best-grownon the Helicobacter agar. To evaluate the total activity of the growing bacteria, ATP assay system wasapplied. The bacterial activities of the cells collected from Blood agar and Modified Skirrow agar after 5days culture were dramatically decreased, whereas the cells from the Helicobacter agar were still increasedor slightly decreased. In addition, the cells collected from the Helicobacter agar after 5 days culturesustained spiral form, while the majority cells from Blood agar and Modified Skirrow agar changed theform into coccoid shape. From these data, we finally concluded that Helicobacter agar (Nissui) was usefulfor rapid detection and also sustaining the bacterial activity.

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