This article was downloaded by: [Columbia University] On: 10 December 2014, At: 09:44 Publisher: Taylor & Francis Informa Ltd Registered in England and Wales Registered Number: 1072954 Registered office: Mortimer House, 37-41 Mortimer Street, London W1T 3JH, UK
Journal of Immunoassay Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/ljii19
Rapid Identification of Candida Albicans by Dot-Enzyme Immunoassay a
a
Roland Guinet , Sylvie Bruneau & Hermance Marlier
a
a
Centre d'Immunochimie Microbienne, Institut Pasteur , Domaine du Poirier, 69210, Lentilly, France Published online: 23 Oct 2006.
To cite this article: Roland Guinet , Sylvie Bruneau & Hermance Marlier (1991) Rapid Identification of Candida Albicans by Dot-Enzyme Immunoassay, Journal of Immunoassay, 12:2, 225-231, DOI: 10.1080/01971529108055068 To link to this article: http://dx.doi.org/10.1080/01971529108055068
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content.
Downloaded by [Columbia University] at 09:44 10 December 2014
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/terms-and-conditions
JOURNAL OF IMMUNOASSAY, 12(2), 225-231 (1991)
Downloaded by [Columbia University] at 09:44 10 December 2014
RAPID IDENTIFICATION OF CANDIDA ALBICANS BY DOT-ENZYM E MMUNOASSAY
Hcland Cuinet, Sylvie Bruneau and Herrnance Marlier C e n t r e d'lmrnuncchimie Micrcbienne, Insti t u t P a s t e u r , D c m a i n e du Pcirier, 69210 Lentilly, F r a n c e
ABSTRACT A highly sensitive and specific d c t - e n z y m e irnrnuncassay f c r t h e rapid identificaticn cf Candida albicans w a s developed using a murine m c n c c l c n a l a n t i b c d y (M a b ) , w h i c h a d s c r b e d t c cell s u r f a c e - e x p c s e d determinants. This Mab r e a c t e d with 28 of t h e 28 C. albicans s t r a i n s t e s t e d including t h e serotypes A and B and 2 C. stellatoidea. I t did n e t r e a c t with 32 c t h e r i s c l a t e s representing eight cther Candida species c e m m o n l y e n c o u n t e r e d in h u m a n m a t e r i a l s . All t h e test c c u l d b e p e r f o r m e d in four s t e p s in less than a n hour. The y e a s t s w e r e directly s p o t t e d on a strip cf Immuncdyne membrane. Then t h e strip w a s incub a t e d f c r 5 min. with t h e Mab, f c r 15 min. with a percxyddse-ccnjugate and for 30 min. w i t h t h e e n z y m e s u b s t r a t e and 4-chlcrc 1 naphtcl. This test prcved useful fcr rapid and easy identificaticn cf C. albicans. (KEY WORDS : Candida albicans, Dct-enzyme irnrnuncassay)
INTRODUCTION Infection d u e t o fungi a r e cf increasing i m p o r t a n c e in p a t i e n t s with c c m p r c m i s e d host d e f e n s e s and t h e m c r t a l i t y r a t e asscciated with t h e s e infections is very high. C. albicans and C. tropicalis a r e t h e species
mcst c c m m c n l y isclated (1). T h e rnicrobiclogical diagncsis, based upcn biochemical tests, is still t i m e consuming (2) in spite cf t h e develcprnent of miniaturized k i t s (3, 4) and of a u t c m a t e d system (5, 6). Slide agglutination tests with specie,-specific 225 Copyright 0 1991 by Marcel Dekker, Inc.
antisera w e r e limited by t h e
226
GUINET, BRUNEAU, AND MARLIER
lack cf specificity cf s e r a since C. albicans s e r c t y p e A and C. tropi-
calis could not b e distinguished (7, 8). A c c m m c n l y used test f c r C. albicans, t h e "germ tube" test, required t h r e e hours and cccasicnal isolates may g i v e a f a l s e negative test. Thus t h e laboratcry n e e d s a fast and sensitive technique tc rapidly
Downloaded by [Columbia University] at 09:44 10 December 2014
identify C. albicans. We r e p o r t h e r e t h e use cf a mcnoclcnal antibody in a d c t - e n z y m e irnrnuncassay f c r t h e rapid d e t e r t i c n cf this microcrganism. MATERIALS A N D M ETHOD5 Y e a s t s strains A t c t a l cf 60 y e a s t s cr yeast-like fungi belcnging t c t h e g e n e r a
Candida,
.%CChWOmyCeS
and Geotrichurn w e r e examined (Table I). M cst
strains h a v e been isolated f r c m pathclcgicdl p r c d u c t s cn Sabcuraud Chlcrampheniccl Agar (Diagncstics P a s t c u r , M a r n e L a C c q u e t t e , France). They w e r e identified by classical c r i t e r i a dcccrding t c van d c r Walt and Yarrow (2). T h e e t h e r strains w e r e stcck c u l t u r e s cbtained f r c m Centraalburedu vcor Schimmelcultures (CBS, Delft, The N e t h e r Idnds), f r c m Naticnal I n s t i t u t e cf Health (NIH, Bethesda, USA) or f r o m l n s t i t u t P a s t e u r (IP, Paris, France). F o r all studies y e a s t s w e r e cultured en Sabcuraud Chlcrarnphenicol Agar f c r 24 H a t 30". D o t e n z y m e irnmuncassay All t h e s t e p s w e r e perf orrned at rocm temperature. S t e p I : T h e blastospores or t h e a r t h r c s p c r e s w e r e remcved f r c m t h e s u r f a c e of t h e a g a r p l a t e and directly s p c t t e d on Imrnuncdyne m e m b r a n e (Pall Industries, St-Germain-en-Layc,
France).
Ten srrains
RAPID IDENTIFICATION OF CANDIDA ALBICANS
227
Downloaded by [Columbia University] at 09:44 10 December 2014
Figure 1 24 H cld c u l t u r e s frcrn : Candida albicans (Ca), C. guilliermondii (Cg), C. krusei (Ck), C. parapSi1osi.s (Cp), C. pseudotropicalis (Cps), C. stellatoidea (Cst), C. tropicalis (C t) and Saccharomyces cerevisiae
(Sc) w e r e directly s p c t t e d en Immuncdyne membrane. A f t e r incubaticn with t h e Mab, t h e percxydase c c n j u g a t e and substrate, s p c t s f r c m C. albicans w e r e s t r c n g l y b l u e c c l c r e d . A l l c t h e r s p c t s w e r e n e t d e tee ted
.
w e r e spotted cn a strip cf 5 x 100 mm. Then t h e strips w e r e scaked in
Tris buffered saline pH 7.5 (TBS : Tris 20 r n M, NaCl 500 rnM) ccntaining 3 % (V/V) gelatin (Bicrad Labcratcries, Richmrnd, USA) f c r 1 min,
t c block t h e remaining sites. S t e p 2 : Incubaticn fcr 5 rnin with t h e Mab
N A B 806,
Cherniccn, El Segundc, Califcrnia, USA) diluted 1 : 200 in TBS ccntaining 0.1 % Tween 20 (TBST, Merck, Darrnstadt, FRC) fcllcwed by twc! washing in TBST f c r I rnin. each. S t e p 3 : Incubaticn f c r 15 min with percxydase ccnjugated rabbit anti-rncuse Ig (Ref. P 161, Dakcpetts, Copenhagen, Denmark) diluted 1 : 200 in TBST. S t e p 4 : A f t e r t h r e e washing in TBST f c r I rnin. and c n e in TBS f c r I min. t h e b l c t s w e r e revealed fcr 30 rnin in sclution cf TRS containing 0.33 % H202(Merrk, Darmstadt, FRG) and 0.05 % 4-chlcrc-1 naphtcl (Sigma C h i m i e SA, L a Verpilliere, France).
GUINET, BRUNEAU, AND MARLIER
228
Table 1 Results of y e a s t s identification in Dct-enzyme immuncassay
Number
Pcsitive results
Negative results
26
26
0
C. albicans received as C . stellatoidea CBS 1905, CRS 2990
2
2
C . glabrata IP 810, 1P 811, 5090, 2036
4
0
4
C. guillierrnondii IP 624, 206
2
0
2
C. krusei 573 CBS, 208 1P
2
0
2
rnacedoniensis CBS 600
1
0
1
3
0
3
3
0
3
5025, 9763, 4620, 5476, 1079, 1559, 4881, 4439, 1325, 1831
13
0
13
S. cerevisiae 1P 605, IP 635, 1248
3
0
3
G. candidurn 1065
I
0
1
Downloaded by [Columbia University] at 09:44 10 December 2014
Tested strains
C . albicans 792 NIH, 311 NIH, 1431, 1422, 1543, 1189, 1513, 184, 34,
HM, 1565, 1434, 1583, 5346, 1285, DUR, 1468, 1485, 214, 243, 740, GIN, BAY, 1309, MOR, BEK
C.
C. parapsilosis IP 45, IP 83, 2147 C.
pseudotropicalis CBS 607, IP 42,
1020
C. tropicalis CBS 94, IP 43, 104,
RAPID IDENTIFICATION OF CANDIDA ALBICANS
229
RESULTST c evaluate the reactivity cf t h e Mab with C . albicans w e tested a panel cf fungi frequently encountered in human materials. A strcng blue cclcur ccrrespcnded t c a pcsitive reacticn easy to read (Figure I). All t h e 28 C. albicans strains were detected including the t w c serctypes A
Downloaded by [Columbia University] at 09:44 10 December 2014
and B and the sucrcse negative variant C. stellatoidea. Ncne cf the cther 32 yeast strains reacted with t h e Mab even C. tropicalis which shared many prcperties with C. QlbiCanS (Table 1). All t h e test cculd be p e r f u m e d i n less than an hour. DISCUSSION
A tctal cf 60 yeasts or yeast-like strains belonging t c the genera Candida, Saccharomyces and Geotrichum w e r e examined. They represented the 8 Candida species the rncst cften isclated frcm human clinical materials and kncwn t c be virulent species. G. candidum and S. cerevisiae w e r e f r e q u e n t l y e n c c u n t e r e d i n c l i n i c a l m a t e r i a l a s
ccmmensal. Using t h e dct-enzyme immunoassay w e cbtained 100 % specificity and 100 % sensitivity fcr the identificaticn cf C. albicans. Ncne cf t h e c t h e r C. albicans i d e n t i f i c a t i c n m e t h c d a c t u a l l y used (biochemical, serclegical or mcrphclcgical) showed such a specificity and sensitivity. Particularly, nc false pcsitive results were cbserved with C. tropicalis. C. albicans and C. tropicalis a r e 2 related species shewing identical DNA C + C % contents (91, and mcre than 80 % of cross-reacting anti-
gens in quantitative immuncelectrcphoresis (10). This test is rapid t c perfcrm : 60 min., versus 3 h for the germtube test, the fastest identification methcd currently available. An
230
GUINET, BRUNEAU, AND MARLIER
i m p o r t a n t point is t h e y e a s t fixaticn on t h e lmmuncdyne strip which is quick and strcng f c r all strains studied. The p r c t c c c l cculd b e shortened by using a n e n z y m e labeled Mab. This test is easy t c perform and results a r e r e a d by visual cbservaticn, which eliminates t h e need for an ELlSA p l a t e reader or a flucres-
Downloaded by [Columbia University] at 09:44 10 December 2014
c e n t rnicrcscope. The develcpment of rapid, sensitive techniques such a s this for t h e d e t e c t i o n of C. albicans infecticns will result in earlier t r e a t m e n t . In additicn this Dct-ELISA test has t h e potential cf being used f c r d i r e c t C. albicans d e t e c t i o n i n clinical swabs.
K EF ER E N C ES 1.
Odds, F.C. Ecclclgy and epidemiclcgy cf Candida. In: Odds, F.C., ed. Candida and Candidcsis. Leicester: Leicester University Press, I9 79: 50-75.
2.
Van d e r Walt, J.P. and Yarrcw, D. Methcds f c r t h e isolation, maintenance, classification and identificaticn cf yeasts. In: Kreger-Van Rij, N.J.W., ed. The yeasts. Amsterdam: Elsevier S c i e n c e Publishers, 1984: 45- 104.
3.
Land, G.A., Harriscn, B.A., Hulme, K.L., Cocper, R.H. and Byrd, J.C. Evaluation cf t h e new API 20C strip for y e a s t identificaticn against a ccnventicnal methcd. J. Clin. Micrcbicl. 1979,lO: 3 5 7 64.
4.
Salkin, I.F., Land, G.A., Hurd, N.J., Goldscn, P.G. and M c Ginnis, M.R. Evaluation cf y e a s t identification and Uni-Yeast-Tek y e a s t i d e n t i f i c a t i c n s y s t e m s . J. C l i n . i 3 i c r c b i c l . 1 9 8 5 2 5 : 6 2 4 - 2 7.
5.
Land, G., S t c t l e r , R., Land, K. and Ztanck, J. U p d a t e and evaluaticn cf t h e a u t c m i c r c b i c y e a s t identificaticn system. J. Clin. Micrcbicl. 1984;20: 649-52.
6. Salkin, I.F., Schadcw, K.H., Bankaitis, L.E., M c Ginnis, M.R. and Kemrna, M.E. Evaluaticn of A b b c t t Quantum 11 y e a s t identificaticn system. J. Clin. Micrcbicl. 1985;22 442-44. 7.
Taguchi, M., Tsubigi, M. and Tsuchiya, T. Rapid identificaticn of y e a s t s by serological methcds. A combined serclcgical and biclcgical methcd. Sabcuraudia 1 9 7 9 ; l Z 185-91.
RAPID IDENTIFICATION OF CANDIDA ALBICANS
231
8. Shincda, T., Kaufman, L. and Padhye, A.H. C o m p a r a t i v e evaluaticn cf t h e i a t r c n sercicgical Candida c h e c k k i t and t h e API 20C k i t i c r t h e i d e n t i f i c a t i c n cf m e d i c a l l y i m p c r t a n t Candida s p e c i e s . 3. Clin. Micrcbicl. 1981;Ik 513-18.
Downloaded by [Columbia University] at 09:44 10 December 2014
9.
Nakase, T. and Kcrnagata, K. Significance cf DNA base ccrnpcsiticn in t h e classificaticn cf y e a s t genus Candida. J. Gen. Appl. illicrcbicl. 1971;l 2 259-79.
10. Gabriel-Bruneau, S.M. and Guinet, K.M .F. Antigenic relaticnships amcng scrne Candida species studied by c,rcssed-line immuncelectrcphcresis : taxcncmic significance. Int. J. Syst. Dactericl. 1984;34: 227-36.
Journal of Immunoassay Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/ljii19
Rapid Identification of Candida Albicans by Dot-Enzyme Immunoassay a
a
Roland Guinet , Sylvie Bruneau & Hermance Marlier
a
a
Centre d'Immunochimie Microbienne, Institut Pasteur , Domaine du Poirier, 69210, Lentilly, France Published online: 23 Oct 2006.
To cite this article: Roland Guinet , Sylvie Bruneau & Hermance Marlier (1991) Rapid Identification of Candida Albicans by Dot-Enzyme Immunoassay, Journal of Immunoassay, 12:2, 225-231, DOI: 10.1080/01971529108055068 To link to this article: http://dx.doi.org/10.1080/01971529108055068
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content.
Downloaded by [Columbia University] at 09:44 10 December 2014
This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms & Conditions of access and use can be found at http://www.tandfonline.com/page/terms-and-conditions
JOURNAL OF IMMUNOASSAY, 12(2), 225-231 (1991)
Downloaded by [Columbia University] at 09:44 10 December 2014
RAPID IDENTIFICATION OF CANDIDA ALBICANS BY DOT-ENZYM E MMUNOASSAY
Hcland Cuinet, Sylvie Bruneau and Herrnance Marlier C e n t r e d'lmrnuncchimie Micrcbienne, Insti t u t P a s t e u r , D c m a i n e du Pcirier, 69210 Lentilly, F r a n c e
ABSTRACT A highly sensitive and specific d c t - e n z y m e irnrnuncassay f c r t h e rapid identificaticn cf Candida albicans w a s developed using a murine m c n c c l c n a l a n t i b c d y (M a b ) , w h i c h a d s c r b e d t c cell s u r f a c e - e x p c s e d determinants. This Mab r e a c t e d with 28 of t h e 28 C. albicans s t r a i n s t e s t e d including t h e serotypes A and B and 2 C. stellatoidea. I t did n e t r e a c t with 32 c t h e r i s c l a t e s representing eight cther Candida species c e m m o n l y e n c o u n t e r e d in h u m a n m a t e r i a l s . All t h e test c c u l d b e p e r f o r m e d in four s t e p s in less than a n hour. The y e a s t s w e r e directly s p o t t e d on a strip cf Immuncdyne membrane. Then t h e strip w a s incub a t e d f c r 5 min. with t h e Mab, f c r 15 min. with a percxyddse-ccnjugate and for 30 min. w i t h t h e e n z y m e s u b s t r a t e and 4-chlcrc 1 naphtcl. This test prcved useful fcr rapid and easy identificaticn cf C. albicans. (KEY WORDS : Candida albicans, Dct-enzyme irnrnuncassay)
INTRODUCTION Infection d u e t o fungi a r e cf increasing i m p o r t a n c e in p a t i e n t s with c c m p r c m i s e d host d e f e n s e s and t h e m c r t a l i t y r a t e asscciated with t h e s e infections is very high. C. albicans and C. tropicalis a r e t h e species
mcst c c m m c n l y isclated (1). T h e rnicrobiclogical diagncsis, based upcn biochemical tests, is still t i m e consuming (2) in spite cf t h e develcprnent of miniaturized k i t s (3, 4) and of a u t c m a t e d system (5, 6). Slide agglutination tests with specie,-specific 225 Copyright 0 1991 by Marcel Dekker, Inc.
antisera w e r e limited by t h e
226
GUINET, BRUNEAU, AND MARLIER
lack cf specificity cf s e r a since C. albicans s e r c t y p e A and C. tropi-
calis could not b e distinguished (7, 8). A c c m m c n l y used test f c r C. albicans, t h e "germ tube" test, required t h r e e hours and cccasicnal isolates may g i v e a f a l s e negative test. Thus t h e laboratcry n e e d s a fast and sensitive technique tc rapidly
Downloaded by [Columbia University] at 09:44 10 December 2014
identify C. albicans. We r e p o r t h e r e t h e use cf a mcnoclcnal antibody in a d c t - e n z y m e irnrnuncassay f c r t h e rapid d e t e r t i c n cf this microcrganism. MATERIALS A N D M ETHOD5 Y e a s t s strains A t c t a l cf 60 y e a s t s cr yeast-like fungi belcnging t c t h e g e n e r a
Candida,
.%CChWOmyCeS
and Geotrichurn w e r e examined (Table I). M cst
strains h a v e been isolated f r c m pathclcgicdl p r c d u c t s cn Sabcuraud Chlcrampheniccl Agar (Diagncstics P a s t c u r , M a r n e L a C c q u e t t e , France). They w e r e identified by classical c r i t e r i a dcccrding t c van d c r Walt and Yarrow (2). T h e e t h e r strains w e r e stcck c u l t u r e s cbtained f r c m Centraalburedu vcor Schimmelcultures (CBS, Delft, The N e t h e r Idnds), f r c m Naticnal I n s t i t u t e cf Health (NIH, Bethesda, USA) or f r o m l n s t i t u t P a s t e u r (IP, Paris, France). F o r all studies y e a s t s w e r e cultured en Sabcuraud Chlcrarnphenicol Agar f c r 24 H a t 30". D o t e n z y m e irnmuncassay All t h e s t e p s w e r e perf orrned at rocm temperature. S t e p I : T h e blastospores or t h e a r t h r c s p c r e s w e r e remcved f r c m t h e s u r f a c e of t h e a g a r p l a t e and directly s p c t t e d on Imrnuncdyne m e m b r a n e (Pall Industries, St-Germain-en-Layc,
France).
Ten srrains
RAPID IDENTIFICATION OF CANDIDA ALBICANS
227
Downloaded by [Columbia University] at 09:44 10 December 2014
Figure 1 24 H cld c u l t u r e s frcrn : Candida albicans (Ca), C. guilliermondii (Cg), C. krusei (Ck), C. parapSi1osi.s (Cp), C. pseudotropicalis (Cps), C. stellatoidea (Cst), C. tropicalis (C t) and Saccharomyces cerevisiae
(Sc) w e r e directly s p c t t e d en Immuncdyne membrane. A f t e r incubaticn with t h e Mab, t h e percxydase c c n j u g a t e and substrate, s p c t s f r c m C. albicans w e r e s t r c n g l y b l u e c c l c r e d . A l l c t h e r s p c t s w e r e n e t d e tee ted
.
w e r e spotted cn a strip cf 5 x 100 mm. Then t h e strips w e r e scaked in
Tris buffered saline pH 7.5 (TBS : Tris 20 r n M, NaCl 500 rnM) ccntaining 3 % (V/V) gelatin (Bicrad Labcratcries, Richmrnd, USA) f c r 1 min,
t c block t h e remaining sites. S t e p 2 : Incubaticn fcr 5 rnin with t h e Mab
N A B 806,
Cherniccn, El Segundc, Califcrnia, USA) diluted 1 : 200 in TBS ccntaining 0.1 % Tween 20 (TBST, Merck, Darrnstadt, FRC) fcllcwed by twc! washing in TBST f c r I rnin. each. S t e p 3 : Incubaticn f c r 15 min with percxydase ccnjugated rabbit anti-rncuse Ig (Ref. P 161, Dakcpetts, Copenhagen, Denmark) diluted 1 : 200 in TBST. S t e p 4 : A f t e r t h r e e washing in TBST f c r I rnin. and c n e in TBS f c r I min. t h e b l c t s w e r e revealed fcr 30 rnin in sclution cf TRS containing 0.33 % H202(Merrk, Darmstadt, FRG) and 0.05 % 4-chlcrc-1 naphtcl (Sigma C h i m i e SA, L a Verpilliere, France).
GUINET, BRUNEAU, AND MARLIER
228
Table 1 Results of y e a s t s identification in Dct-enzyme immuncassay
Number
Pcsitive results
Negative results
26
26
0
C. albicans received as C . stellatoidea CBS 1905, CRS 2990
2
2
C . glabrata IP 810, 1P 811, 5090, 2036
4
0
4
C. guillierrnondii IP 624, 206
2
0
2
C. krusei 573 CBS, 208 1P
2
0
2
rnacedoniensis CBS 600
1
0
1
3
0
3
3
0
3
5025, 9763, 4620, 5476, 1079, 1559, 4881, 4439, 1325, 1831
13
0
13
S. cerevisiae 1P 605, IP 635, 1248
3
0
3
G. candidurn 1065
I
0
1
Downloaded by [Columbia University] at 09:44 10 December 2014
Tested strains
C . albicans 792 NIH, 311 NIH, 1431, 1422, 1543, 1189, 1513, 184, 34,
HM, 1565, 1434, 1583, 5346, 1285, DUR, 1468, 1485, 214, 243, 740, GIN, BAY, 1309, MOR, BEK
C.
C. parapsilosis IP 45, IP 83, 2147 C.
pseudotropicalis CBS 607, IP 42,
1020
C. tropicalis CBS 94, IP 43, 104,
RAPID IDENTIFICATION OF CANDIDA ALBICANS
229
RESULTST c evaluate the reactivity cf t h e Mab with C . albicans w e tested a panel cf fungi frequently encountered in human materials. A strcng blue cclcur ccrrespcnded t c a pcsitive reacticn easy to read (Figure I). All t h e 28 C. albicans strains were detected including the t w c serctypes A
Downloaded by [Columbia University] at 09:44 10 December 2014
and B and the sucrcse negative variant C. stellatoidea. Ncne cf the cther 32 yeast strains reacted with t h e Mab even C. tropicalis which shared many prcperties with C. QlbiCanS (Table 1). All t h e test cculd be p e r f u m e d i n less than an hour. DISCUSSION
A tctal cf 60 yeasts or yeast-like strains belonging t c the genera Candida, Saccharomyces and Geotrichum w e r e examined. They represented the 8 Candida species the rncst cften isclated frcm human clinical materials and kncwn t c be virulent species. G. candidum and S. cerevisiae w e r e f r e q u e n t l y e n c c u n t e r e d i n c l i n i c a l m a t e r i a l a s
ccmmensal. Using t h e dct-enzyme immunoassay w e cbtained 100 % specificity and 100 % sensitivity fcr the identificaticn cf C. albicans. Ncne cf t h e c t h e r C. albicans i d e n t i f i c a t i c n m e t h c d a c t u a l l y used (biochemical, serclegical or mcrphclcgical) showed such a specificity and sensitivity. Particularly, nc false pcsitive results were cbserved with C. tropicalis. C. albicans and C. tropicalis a r e 2 related species shewing identical DNA C + C % contents (91, and mcre than 80 % of cross-reacting anti-
gens in quantitative immuncelectrcphoresis (10). This test is rapid t c perfcrm : 60 min., versus 3 h for the germtube test, the fastest identification methcd currently available. An
230
GUINET, BRUNEAU, AND MARLIER
i m p o r t a n t point is t h e y e a s t fixaticn on t h e lmmuncdyne strip which is quick and strcng f c r all strains studied. The p r c t c c c l cculd b e shortened by using a n e n z y m e labeled Mab. This test is easy t c perform and results a r e r e a d by visual cbservaticn, which eliminates t h e need for an ELlSA p l a t e reader or a flucres-
Downloaded by [Columbia University] at 09:44 10 December 2014
c e n t rnicrcscope. The develcpment of rapid, sensitive techniques such a s this for t h e d e t e c t i o n of C. albicans infecticns will result in earlier t r e a t m e n t . In additicn this Dct-ELISA test has t h e potential cf being used f c r d i r e c t C. albicans d e t e c t i o n i n clinical swabs.
K EF ER E N C ES 1.
Odds, F.C. Ecclclgy and epidemiclcgy cf Candida. In: Odds, F.C., ed. Candida and Candidcsis. Leicester: Leicester University Press, I9 79: 50-75.
2.
Van d e r Walt, J.P. and Yarrcw, D. Methcds f c r t h e isolation, maintenance, classification and identificaticn cf yeasts. In: Kreger-Van Rij, N.J.W., ed. The yeasts. Amsterdam: Elsevier S c i e n c e Publishers, 1984: 45- 104.
3.
Land, G.A., Harriscn, B.A., Hulme, K.L., Cocper, R.H. and Byrd, J.C. Evaluation cf t h e new API 20C strip for y e a s t identificaticn against a ccnventicnal methcd. J. Clin. Micrcbicl. 1979,lO: 3 5 7 64.
4.
Salkin, I.F., Land, G.A., Hurd, N.J., Goldscn, P.G. and M c Ginnis, M.R. Evaluation cf y e a s t identification and Uni-Yeast-Tek y e a s t i d e n t i f i c a t i c n s y s t e m s . J. C l i n . i 3 i c r c b i c l . 1 9 8 5 2 5 : 6 2 4 - 2 7.
5.
Land, G., S t c t l e r , R., Land, K. and Ztanck, J. U p d a t e and evaluaticn cf t h e a u t c m i c r c b i c y e a s t identificaticn system. J. Clin. Micrcbicl. 1984;20: 649-52.
6. Salkin, I.F., Schadcw, K.H., Bankaitis, L.E., M c Ginnis, M.R. and Kemrna, M.E. Evaluaticn of A b b c t t Quantum 11 y e a s t identificaticn system. J. Clin. Micrcbicl. 1985;22 442-44. 7.
Taguchi, M., Tsubigi, M. and Tsuchiya, T. Rapid identificaticn of y e a s t s by serological methcds. A combined serclcgical and biclcgical methcd. Sabcuraudia 1 9 7 9 ; l Z 185-91.
RAPID IDENTIFICATION OF CANDIDA ALBICANS
231
8. Shincda, T., Kaufman, L. and Padhye, A.H. C o m p a r a t i v e evaluaticn cf t h e i a t r c n sercicgical Candida c h e c k k i t and t h e API 20C k i t i c r t h e i d e n t i f i c a t i c n cf m e d i c a l l y i m p c r t a n t Candida s p e c i e s . 3. Clin. Micrcbicl. 1981;Ik 513-18.
Downloaded by [Columbia University] at 09:44 10 December 2014
9.
Nakase, T. and Kcrnagata, K. Significance cf DNA base ccrnpcsiticn in t h e classificaticn cf y e a s t genus Candida. J. Gen. Appl. illicrcbicl. 1971;l 2 259-79.
10. Gabriel-Bruneau, S.M. and Guinet, K.M .F. Antigenic relaticnships amcng scrne Candida species studied by c,rcssed-line immuncelectrcphcresis : taxcncmic significance. Int. J. Syst. Dactericl. 1984;34: 227-36.
