酵素抗体法（迅速キット）によるClostridiumdifficileトキシン …

1 2013.. Clostridium difficile . C. difficile . 1 1 1 1 1 1 . 1 1 2 2 2 1 3 . 1 .. 2 .. 3 .. 24 11 6 25 10 24 . Clostridium difficile A B TOX A!B QUIK CHEK. TOX A! B C. difficile . toxin B C. difficile . C. difficile Clostridium difficile infection CDI . 232 92 92 PCR toxin A toxin B toxin A toxin B 55 toxin A. toxin B 11 toxin A toxin B 26 toxin B 66 Toxin B C. difficile . TOX A! B 28! 66 164! 166 .. TOX A! B . CDI C. difficile glutamate dehydrogenase .. Key words: Clostridium difficile TOX A! B QUIK CHEK . CDI . Clostridium difficile 5 10 C. difficile A B 2 .. C. difficile gold Clostridium difficile infection standard . CDI CDI 48 . antibiotic-associated diarrhea AAD 15.

2 25 C. difficile . C. difficile A B . TOX A! B QUIK CHEK . 113-8431 3-1-3. toxin B C. difficile .. TEL: 03-3813-3111 5172 . Vol. 23 No. 4 2013. 7. 260 . Table 1. TOX A/B QUIK CHEK . toxin B C. difficile . 2007 9 2010 8 . C. difficile . 355 Toxin B . CDI .. 28 1 1. 3 TOX A/B. 38 25 139. 2 . 66 26 140.. A B 55 A B 11 A B 26 .. 232 . C. difficile enzyme im- 120 annealing 35 . munoassay EIA TOX A! B QUIK CHEK . TECHLAB ! TOX A! PCR. B TOX A! B A B toxin A toxin B . TOX A B toxin A toxin B A B toxin A!B A toxin B A B . 2 Toxin B C. difficile . 4 80 . Fisher . C. difficile CCMA EX 2 . CCMA Stat Flex version 35 48 p . C. difficile CCMA .. 103! ml 1 104 . 105!

3 Ml 2 106! ml 3 1 TOX A! B QUIK CHEK . toxin A toxin B toxin B C. difficile . PCR 232 TOX A! B C. 50 l McFar- difficile toxin B . land No. 1 100 10 Table 1 C. difficile 232 . l template DNA 92 92 toxin PCR TAKARA TAQ R001B B C. difficile 66 A B 55 A B 11. 25 l 28 . Gene AmpPCR System 9600-R Toxin B C. difficile 26 . Toxin A NK9 1 . 5 -CCA CCA GCT GCA GCC ATA-3 NK11 5 -TGA 140 1 . TGC TAA TAA TGA ATC TAA AAT GGT AAC- TOX A! B . 3 NKV011 5 -TTT TGA TCC TAT AGA 28! 66 . 164! 166 . ATC TAA CTT AGT AAC-3 95 20 . denature 62 120 annealing 35 2 Toxin B C. difficile . Toxin B NK104 . 5 -GTG TAG CAA TGA AAG TCC AAG TTT ACG Toxin B C. difficile 66 . C-3 . NK105 5 -CAC TTA GCT CTT TGA TTG CTG.

4 CAC CT-3 95 20 denature 55 Table 2 . 8 Vol. 23 No. 4 2013. C. difficile 261. Table 2. Toxin B C. difficile .. TOX A/B QUIK CHEK Odds . a p b 95% . 1 5 14 1. 2 5 10 . 3 18 14 . a 1 103/ml 2 104 105/ml 3 106/ml . b 1 .. Fisher 2 . 1 2 3 . 3 1 TOX A! B . p . EIA . CDI .. Toxin B C. difficile . TOX A! B . CDI. TOX A! B GDH GDH .. gold standard . 20 . C. difficile 2009 1 . TOX A! B .. 1 Kato, H., H. Kita, T. Karasawa, et al. 2001. Colonisation and transmission of Clostridium difficile in healthy individuals examined by PCR ribotyping and pulsed- Shin field gel electrophoresis. J. Med. Microbiol. 50: 720- CDI 727. 2 Cohen, , Gerding, S. Johnson, et al. 2010.

5 Clini- cal practice guidelines for Clostridium difficile infec- CDI . tion in adults: 2010 update by the Society for Health- Goldenberg . care Epidemiology of America (SHEA) and the Infec- EIA . tious Diseases Society of America (IDSA). Infect. Con- C. difficile gluta- trol Hosp. Epidemiol. 31: 431-455. mate dehydrogenase GDH . 3 2008 . TOX A! B . Clostridium difficile . GDH . 82: 419-426. TOX A! B . Vol. 23 No. 4 2013. 9. 262 . 4 Kuijper, , J. de Weerdt, H. Kato, et al. 2001. Noso- 12 Eastwood, K., P. Else, A. Charlett, et al. 2009. Com- comial outbreak of Clostridium difficile -associated di- parison of nine commercially available Clostridium arrhoea due to a clindamycin-resistant enterotoxin difficile toxin detection assays, a real-time PCR assay A-negative strain.

6 Eur. J. Clin. Microbiol. Infect. Dis. for C. difficile tcdB , and a glutamate dehydrogenase 20: 528-534. detection assay to cytotoxin testing and cytotoxi- 5 2002 Clostridium difficile genic culture methods. J. Clin. Microbiol. 47: 3211- 12: 115-122. 3217. 6 Limaye, , Turgeon, Cookson, et al. 2000. 13 2011 . Pseudomembranous colitis caused by a toxin A B Clostridium difficile Toxin A Toxin B . strain of Clostridium difficile . J. Clin. Microbiol. 21: 51- 38: 1696-1697. 58. 7 Lyras, D., O Connor, Howarth, et al. 2009. 14 2009 . Toxin B is essential for virulence of Clostridium diffi- TOX A!B QUIKCHEK . cile . Nature 458: 1176-1179. 58: 938-942. 8 Kato, H.

7 , N. Kato, K. Watanabe, et al. 1998. Identifica- 15 2011 Clostrid- tion of toxin A-negative, toxin B-positive Clostridium ium difficile C. dif- difficile by PCR. J. Clin. Microbiol. 36: 2178-2182. ficile A!B C. 9 Kato, H., N. Kato, S. Katow, et al. 1999. Deletions in DIFF QUIK CHEK COMPLETE . the repeating sequences of the toxin A gene of toxin 21: 253-259. A-negative, toxin B-positive Clostridium difficile 16 Shin, , Kuak, Lee, et al. 2009. Algorithm strains. FEMS Microbiol. Lett. 175: 197-203. combining toxin immunoassay and stool culture for 10 Turgeon, , Novicki, J. Quick, et al. 2003. Six diagnosis of Clostridium difficile infection. J. Clin. Mi- rapid tests for direct detection of Clostridium difficile crobiol.

8 47: 2952-2956. and its toxins in fecal samples compared with the fi- 17 Goldenberg, , Cliff, French. 2010. Labora- broblast cytotoxicity assay. J. Clin. Microbiol. 41: 667- tory diagnosis of Clostridium difficile infection. J. 670. Clin. Microbiol. 48: 3048-3049. 11 Alcal , L., L. S nchez-Cambronero, Catal n, et al. 18 2008 Clostrid- 2008. Comparison of three commercial methods for ium difficile toxin A toxin rapid detection of Clostridium difficile toxins A and B TOX A!B QUIK CHEK . B from fecal specimens. J. Clin. Microbiol. 46: 3833- 38: 52-54. 3835. 10 Vol. 23 No. 4 2013. C. difficile 263. Evaluation of a commercial enzyme immunoassay test kit for detection of Clostridium difficile toxins A and B in stool specimens Mayumi Oki 1) , Shigeki Misawa 1) , Hiromi Arai 1) , Yukiko Kawase 1) , Masayoshi Chounan 1) , Takaaki Kawakami 1) , Yutaka Kuno 1) , Takashi Horii 1) , Shigemi Kondo 2) , Kazunori Miyake 2) , Yoko Tabe 2) , Akimichi Ohsaka 1) 3).

9 1). Department of Clinical Laboratory, Juntendo University Hospital 2). Department of Clinical Laboratory Medicine, Juntendo University School of Medicine 3). Department of Transfusion Medicine and Stem Cell Regulation, Juntendo University School of Medicine We evaluated the performance of detection of Clostridium difficile toxins A and B in stool specimens by a rapid enzyme immunoassay test kit, TOX A! B QUIK CHEK (TOX A! B), in comparison with toxigenic culture as the reference method. In addition, semiquantitative analysis of C. difficile culture was compared with the stool toxin detection by TOX A! B. A total of 232 diarrheal stool specimens were collected and assayed from patients sus- pected of having C.

10 Difficile infection (CDI). Of the 92 culture-positive specimens, PCR analysis revealed 55 toxin A- and toxin B-positive isolates, 11 toxin A-negative and toxin B-positive isolates, and 26 toxins A- and B-negative isolates. Therefore, 66 specimens showed positive toxigenic culture results; in comparison with the toxigenic cul- ture method, the sensitivity and specificity of the TOX A! B test for detection of toxins A and B in stool specimens were (28! 66) and (164! 166), respectively. The results of toxin detection by TOX A! B were likely posi- tive in the specimen with the greater number of colonies on semiquantitative culture plate. Because the TOX A!