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Bacteriology

Molecular Serotyping of Klebsiella Species Isolates by Restriction of the Amplified Capsular Antigen Gene Cluster

Sylvain Brisse, Sylvie Issenhuth-Jeanjean, Patrick A. D. Grimont
Sylvain Brisse
Unité Biodiversité des Bactéries Pathogènes Emergentes (U389 INSERM), Institut Pasteur, Paris, France
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  • For correspondence: sbrisse@pasteur.fr
Sylvie Issenhuth-Jeanjean
Unité Biodiversité des Bactéries Pathogènes Emergentes (U389 INSERM), Institut Pasteur, Paris, France
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Patrick A. D. Grimont
Unité Biodiversité des Bactéries Pathogènes Emergentes (U389 INSERM), Institut Pasteur, Paris, France
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DOI: 10.1128/JCM.42.8.3388-3398.2004
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  • FIG. 1.
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    FIG. 1.

    cps PCR-RFLP analysis of 12 Klebsiella strains. Lanes M, molecular size markers (in base pairs). The C-pattern codes are indicated above the lanes, and the K serotypes and strain names are given in parentheses. Small differences can be observed between patterns C2d and C2e, on the one hand, and between patterns C4a, C4b, and C4c, on the other. C pattern C4f shares only two fragments with the other C patterns of K4 strains.

  • FIG. 2.
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    FIG. 2.

    Overview of the 102 distinct C patterns found during this study. The five C patterns obtained for clinical strains whose K serotypes were initially unknown and which did not exactly match a reference C pattern are given. The patterns obtained with primer rCPS2 (Fig. 3) are not included. C patterns were ordered by similarity by the unweighted pair group method with arithmetic averages procedure, based on the Pearson coefficient, by using a 1% optimization parameter. Pearson coefficients were converted to percentages with BioNumerics software. Uncertain bands, which were weak and which were not taken into account for pattern comparison, are indicated by dashes.

  • FIG. 3.
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    FIG. 3.

    Comparison of the patterns obtained after amplification with the alternative primer, gnd-specific primer rCPS2, with closely matching C patterns obtained with primer rCPS (and BioNumerics software). In most cases, the patterns obtained with primer rCPS2 differed by one or two bands from the corresponding C pattern obtained for strains of the same K serotype. The only exceptions are for K68 strain 1578 and K6 strain 1283; the C pattern of the latter strain nevertheless shares four fragments with pattern C6a.

Tables

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  • TABLE 1.

    Overview of the 246 strains evaluated in this study

    Strain(s)aK serotype(s)bC patternSpeciescSource 1eSource 2fDate(s)
    Strains
        SB165, CIP 52.2081C1a K. p. subsp. pneumoniaeK1 ref. strainA 5054
        CIP A2151C1a K. p. subsp. pneumoniaeEyelidFrance, France1954
        CIP 52.1441C1a K. p. subsp. pneumoniaeLaboratory strain
        3981C1a K. p. subsp. pneumoniaeSputumLouvain, Belgium1976
        4081C1a K. p. subsp. pneumoniaeBloodParis, France1976
        4281C1a K. p. subsp. pneumoniaeBloodSt Etienne, France1976
        4401C1a K. p. subsp. pneumoniaeBloodParis, France1977
        4671C1a K. p. subsp. pneumoniaeLaboratory strainLyon, France1977
        4681C1a K. p. subsp. pneumoniaeTracheaParis, France1977
        5031C1a K. p. subsp. pneumoniaeSputumRoanne, France1977
        6431C1a K. p. subsp. pneumoniaeCSFParis, France1980
        6871C1a K. p. subsp. pneumoniaeCSFCréteil, France1980
        7441C1a K. p. subsp. pneumoniaeMare, metritisParis, France1981
        7981C1a K. p. subsp. pneumoniaeSputumParis, France1982
        9201C1a K. p. subsp. pneumoniaePleural fluidPontarlier, France1984
        9261C1a K. p. subsp. pneumoniaeBloodParis, France1984
        SB166, CIP 52.1452C2a K. p. subsp. pneumoniaeK2 ref. strainB 5055
        6752C2a K. p. subsp. pneumoniaeVaccine strainMontpellier, France1980
        15942C2a K. p. subsp. pneumoniaeMonkeyMulhouse, France1988
        15842C2b K. p. subsp. pneumoniaeBloodBriançon, France1988
        7772C2c K. p. subsp. pneumoniaeBloodParis, France1981
        342C2d K. p. subsp. pneumoniaeLaboratory strain
        CIP 52.2042C2e K. p. subsp. pneumoniaeLaboratory strain
        352C2e K. p. subsp. pneumoniaeLaboratory strain
        7782C2e K. p. subsp. pneumoniaeCSFParis, France1981
        7792C2e K. p. subsp. pneumoniaeAbcessParis, France1981
        SB167, CIP 52.1463C3a K. p. subsp. pneumoniaeK3 ref. strainC 5046
        CIP 52.210T, ATCC 13884T3C3a K. p. subsp. rhinoscleromatisType strain
        CIP 82.91T, ATCC 13883T3C3a K. p. subsp. pneumoniaeType strain
        ATCC 69083C3a K. p. subsp. rhinoscleromatisReference strain
        CIP 54.453C3a K. p. subsp. rhinoscleromatisNoseVietnam1954
        CIP 54.1293C3a K. p. subsp. rhinoscleromatisRhinoscleromaVietnam1954
        4753C3a K. p. subsp. rhinoscleromatisSputumPoland1977
        5383C3a K. p. subsp. pneumoniaeUrineParis, France1977
        7453C3a K. p. subsp. rhinoscleromatisGranulomaAlgérie1981
        7463C3a K. p. subsp. rhinoscleromatisRhinoscleromaAlgeria1981
        8393C3a K. p. subsp. rhinoscleromatisNoseParis, France1982
        15093C3a K. p. subsp. pneumoniaeFecesParis, France1988
        16353C3a K. p. subsp. pneumoniaePusParis, France1989
        16493C3a K. p. subsp. pneumoniaeBloodParis, France1989
        34.813C3a K. p. subsp. rhinoscleromatisNoseFrance1981
        42.793C3a K. p. subsp. rhinoscleromatisNoseParis, France1979
        SB17853C3a K. p. subsp. rhinoscleromatisSputumPoland1977
        SB17933C3a K. p. subsp. rhinoscleromatisGranulomaWien, Austria2001
        SB168, CIP 52.211T4C4a K. p. subsp. ozaenaeK4 ref. strainD 5050
        ATCC 11296 T4C4a K. p. subsp. ozaenaeType strainIndonesia1930
        8634C4a K. p. subsp. ozaenaeSputumLyon, France1983
        15754C4a K. p. subsp. ozaenaeBloodParis, France1988
        12024C4a K. p. subsp. ozaenaeBloodParis, France1986
        15104C4a K. p. subsp. ozaenaeEar pusParis, France1988
        9194C4b K. p. subsp. ozaenaeLungTours, France1989
        9094C4c K. p. subsp. ozaenaeBronchial aspPontoise, France1984
        16114C4d K. p. subsp. ozaenaeBloodCaen, France1989
        7484C4e K. p. subsp. ozaenaeNoseAlgeria1981
        8894C4f K. p. subsp. ozaenaeBronchial aspGrenoble, France1983
        CIP 52.2125C5a K. p. subsp. ozaenaeK5 ref. strainE 5051
        1895C5a K. p. subsp. pneumoniaeLaboratory strain1973
        3255C5a K. p. subsp. pneumoniaeHumanParis, France1975
        4215C5a K. p. subsp. ozaenaeNoseKigali, Rwanda1976
        CIP 52.2136C6a K. p. subsp. pneumoniaeK6 ref. strainF 5052
        CIP 80.426C6b K. p. subsp. pneumoniaeWoodCompiègne, France1979
        SB179, CIP 52.2057C7a K. p. subsp. pneumoniaeK7 ref. strainAerogenes 4140
        15437C7a K. p. subsp. pneumoniaeHorse, penisParis, France1988
        15637C7a K. p. subsp. pneumoniaeHorse, penisParis, France1989
        15837C7a K. p. subsp. pneumoniaeHorseCaen, France1988
        16047C7a K. p. subsp. pneumoniaeHorse, spermParis, France1988
        CIP 52.2068C8a K. planticola K8 ref. strain Klebsiella 1015
        15748C8b K. p. subsp. pneumoniaeBloodParis, France1988
        SB181, CIP 52.2079C9a K. p. subsp. pneumoniaeK9 ref. strain Klebsiella 1056
        SB182, CIP 52.21410C10a K. p. subsp. pneumoniaeK10 ref. strain Klebsiella 919Source 1eSource 2fDate(s)
        114110C10b K. p. subsp. pneumoniaeBronchial aspLyon, France1986
        CIP 52.21511C11a K. p. subsp. pneumoniaeK11 ref. strain Klebsiella 390
        SB184, CIP 52.21612C12a K. p. subsp. pneumoniaeK12 ref. strain Klebsiella 313
        SB185, CIP 52.21713C13a K. p. subsp. pneumoniaeK13 ref. strain Klebsiella 1470
        SB186, CIP 52.21814C14a K. planticola K14 ref. strain138
        SB187, CIP 52.21915C15a K. p. subsp. pneumoniaeK15 ref. strainMich. 61
        SB188, CIP 52.22016C16a K. p. subsp. pneumoniaeK16 ref. strain2069/49
        163216C16a K. p. subsp. pneumoniaeSeptic tankParis, France1988
        SB189, CIP 52.22117C17a K. p. subsp. pneumoniaeK17 ref. strain2005/49
        SB190, CIP 52.22218C18a K. p. subsp. pneumoniaeK18 ref. strain1754/49
        SB191, CIP 52.22319C19a K. p. subsp. pneumoniaeK19 ref. strain293/50
        SB19220C20a K. p. subsp. pneumoniaeK20 ref. strain889/50
        SB193, CIP 52.22521C21a K. p. subsp. pneumoniaeK21 ref. strain1702/49
        SB194, CIP 52.22622C22,37a K. p. subsp. pneumoniaeK22 ref. strain1996/49
        SB195, CIP 52.22823C23a K. p. subsp. pneumoniaeK23 ref. strain2812/50
        SB196, CIP 52.22924C24a K. p. subsp. pneumoniaeK24 ref. strain1680/49
        158924C24a K. p. subsp. pneumoniaeBloodParis, France1988
        SB197, CIP 52.23025C25a K. p. subsp. pneumoniaeK25 ref. strain2002/49
        158125C25b K. p. subsp. pneumoniaeBloodParis, France1988
        160125C25b K. p. subsp. pneumoniaeFecesCréteil, France1988
        SB198, CIP 53.626C26a K. oxytoca K26 ref. strain5884
        SB199, CIP 52.23227C27a K. p. subsp. pneumoniaeK27 ref. strain6613
        SB200, CIP 52.23328C28a K. p. subsp. pneumoniaeK28 ref. strain5758
        SB317, CIP 52.23429C29a K. oxytoca K29 ref. strain5725y
        SB318, CIP 52.23530C30a K. p. subsp. pneumoniaeK30 ref. strain7824
        156930C30a K. p. subsp. pneumoniaeHorse, penisParis, France1988
        162330C30a K. p. subsp. pneumoniaeHorse, spermSaint Lo, France1988
        155330C30b K. p. subsp. pneumoniaeHorse, spermLyon, France1989
        SB319, CIP 52.23131C31a K. p. subsp. pneumoniaeK31 ref. strain6258
        SB33932C32a K. planticola K32 ref. strain6837
        SB473, CIP 53.833C33a K. p. subsp. pneumoniaeK33 ref. strain6168
        SB474, CIP 53.934C34a K. p. subsp. pneumoniaeK34 ref. strain7522
        SB475, CIP 53.1035C35a K. planticola K35 ref. strain7444
        151635C35b K. p. subsp. pneumoniaeUrineParis, France1988
        138635C35c K. p. subsp. pneumoniaeEnterocolitisMontpellier, France1987
        SB476, CIP 53.1136C36a K. p. subsp. pneumoniaeK36 ref. strain8306
        SB477, CIP 53.1237C22,37a K. p. subsp. pneumoniaeK37 ref. strain8238
        CIP 53.1338C38a K. p. subsp. pneumoniaeK38 ref. strain8414
        SB479, CIP 53.1439C39a K. planticola K39 ref. strain7749
        58539C39b K. p. subsp. pneumoniaeHandSt Etienne, France1979
        SB480, CIP 53.1540C40a K. p. subsp. pneumoniaeK40 ref. strain8588
        SB48141C41a K. p. subsp. pneumoniaeK41 ref. strain6177
        SB482, CIP 53.1742C42a K. p. subsp. pneumoniaeK42 ref. strain1702
        SB483, CIP 53.1943C43a K. p. subsp. pneumoniaeK43 ref. strain2482
        SB484, CIP 53.2044C44a K. planticola K44 ref. strain7730
        SB485, CIP 53.2145C45a K. p. subsp. pneumoniaeK45 ref. strain8464
        SB486, CIP 53.2246C46a K. p. subsp. pneumoniaeK46 ref. strain5281
        SB487, CIP 53.2347C47a K. p. subsp. pneumoniaeK47 ref. strain9682
        70047C47a K. p. subsp. pneumoniaeWood, tropicalCompiègne, France1980
        165947C47b K. p. subsp. pneumoniaeFecesParis, France1989
        SB488, CIP 53.2448C48a K. planticola K48 ref. strain1196
        SB489, CIP 52.19949C49a K. planticola K49 ref. strain6115
        CIP 52.20050C50a K. p. subsp. pneumoniaeK50 ref. strain1303/50
        SB491, CIP 52.20151C51a K. p. subsp. pneumoniaeK51 ref. strain4715/50
        SB49252C52a K. p. subsp. pneumoniaeK52 ref. strain5759/50
        MGH 7857852C52a K. p. subsp. pneumoniaeReference strain
        157952C52b K. omithinolytica Bronchial aspToulouse, France1988
        SB493, CIP 53.2653C53a K. planticola K53 ref. strain1756/51
        SB494, CIP 53.2754C54a K. planticola K54 ref. strainStanley
        SB49555C55a K. p. subsp. pneumoniaeK55 ref. strain3985/51
        SB49656C56a K. planticola K56 ref. strain3534/51
        SB49757C57a K. planticola K57 ref. strain4425/51
        SB49858C58a K. planticola K58 ref. strain636/52
        9459C59a K. oxytoca UnknownParis, France1972
        SB50060C60a K. p. subsp. pneumoniaeK60 ref. strain4463/52
        SB50161C61a K. p. subsp. pneumoniaeK61 ref. strain5710/52
        SB50262C62a K. p. subsp. pneumoniaeK62 ref. strain5711/52
        162962C62a K. p. subsp. pneumoniaeDonkeyMulhouse, France1988
        151362C62b K. p. subsp. pneumoniaeFecesParis, France1988
        19962C62c K. oxytoca Bronchial aspParis, France1974
        SB50363C63a K. p. subsp. pneumoniaeK63 ref. strain5845/52
        SB50464C64a K. p. subsp. pneumoniaeK64 ref. strainNCTC 8172
        SB50565C65a K. terrigena K65 ref. strainSW 4
        SB50666C66a K. oxytoca K66 ref. strain438 (3a)
        SB50767C67a K. p. subsp. pneumoniaeK67 ref. strain264 (1)
        118768C68a K. oxytoca WoundParis, France1986
        134468C68a K. oxytoca BloodBordeaux, France1986
        SB50869C69a K. p. subsp. pneumoniaeK69 ref. strain889
        SB50970C70a K. oxytoca K70 ref. strain167
        SB51071C71a K. planticola K71 ref. strain4349
        SB51172C72a K. planticola K72 ref. strain1205
        SB51274C74a K. oxytoca K74 ref. strain371
        SB51379C79a K. planticola K79 ref. strain325
        SB51480C80a K. p. subsp. pneumoniaeK80 ref. strain708
        SB51581C81a K. p. subsp. pneumoniaeK81 ref. strain370
        SB51682C82a K. p. subsp. pneumoniaeK82 ref. strain3454-70
        10371,58C1a K. p. subsp. pneumoniaeHorsePont l’Eveque, France1985
        10561,58C1a K. p. subsp. pneumoniaeHorseCaen, France1985
        15701,58C1a K. p. subsp. pneumoniaeMare, metritisParis, France1988
        60721,31C21,31a K. p. subsp. pneumoniaeUrineSaint-Etienne, France1978
        162222,37C22,37a K. p. subsp. ozaenaeSputumMali1989
        683AcapsularC2e K. p. subsp. pneumoniaeHumanLyon, France1980
        1340AcapsularC3a K. p. subsp. pneumoniaeBloodBordeaux, France1987
        156730C30a K. p. subsp. pneumoniaeHorse, penisParis, France1988
        1609AcapsularC14a K. p. subsp. pneumoniaeUrineParis, France1988
        SB49959No or weak PCR product K. planticola K59 ref. strain2212/52
        15862No or weak PCR product K. p. subsp. pneumoniaeBloodBriançon, France1988
        12836No or weak PCR product K. terrigena CarrotsMulhouse, France1988
        14166No or weak PCR product K. terrigena Horse, spermNantes, France1988
        1476AcapsularNo or weak PCR product K. p. subsp. pneumoniaeHospital env.Rennes, France1988
        148316No or weak PCR product K. p. subsp. pneumoniaeHospital env.Rennes, France1988
        148524No or weak PCR product K. p. subsp. pneumoniaeHospital env.Rennes, France1988
        15447No or weak PCR product K. p. subsp. pneumoniaeHorse, penisParis, France1988
        157868No or weak PCR product K. oxytoca BloodParis, France1988
        13469No unique PCR product K. p. subsp. pneumoniaeBloodBordeaux, France1987
        15476No unique PCR product K. p. subsp. pneumoniaeMare, uterusCaen, France1988
        15763No unique PCR product K. p. subsp. pneumoniaeUrineParis, France1988
        7754No unique PCR product K. p. subsp. ozaenaeNoseDakar, Senegal1981
    Strains in challenge collection of clinical isolatesd
        SB5422, 37C22,37a K. p. subsp. pneumoniaeBloodSeville, Spain1997-1998
        SB5237C22,37a K. p. subsp. pneumoniaeBloodSeville, Spain1997-1998
        SB111124C24a K. p. subsp. pneumoniaeUrineCoimbra, Portugal1997-1998
        SB6425C25b K. p. subsp. pneumoniaeBloodLondon, United Kingdom1997-1998
        SB11827C27a K. p. subsp. pneumoniaeWoundAthens, Greece1997-1998
        SB13827C27a K. p. subsp. pneumoniaeWoundAthens, Greece1997-1998
        SB15330C30a K. p. subsp. pneumoniaeBloodAnkara, Turkey1997-1998
        SB16230C30a K. p. subsp. pneumoniaeBloodIstanbul, Turkey1997-1998
        SB22330C30a K. p. subsp. pneumoniaeBloodAnkara, Turkey1997-1998
        SB11430C30b K. p. subsp. pneumoniaeBloodAthens, Greece1997-1998
        SB59230C30b K. p. subsp. pneumoniaeBloodAthens, Greece1997-1998
        SB9339C39b K. p. subsp. pneumoniaeBloodKrakow, Poland1997-1998
        SB21052C52a K. p. subsp. pneumoniaeUrineLille, France1997-1998
        SB4462C62a K. p. subsp. pneumoniaeBloodUtrecht, The Netherlands1997-1998
        SB16435C-NEW-1 K. p. subsp. pneumoniaeBloodAthens, Greece1997-1998
        SB20422, 37CNEW-2 K. p. subsp. pneumoniaeBloodLondon, United Kingdom1997-1998
        SB120NTCNEW-3 K. p. subsp. pneumoniaeBloodGenoa, Italy1997-1998
        SB39NTCNEW-3 K. p. subsp. pneumoniaeBloodGenoa, Italy1997-1998
        SB11324C24alike-a K. p. subsp. pneumoniaeUrineLille, France1997-1998
        SB15868C68alike-a K. p. subsp. pneumoniaeUrineGenoa, Italy1997-1998
        03-91382C2a K. p. subsp. pneumoniaeAbcessFrance2003
        03-24553C3a K. p. subsp. rhinoscleromatisNoseMali2003
    • ↵ a Entries with two strain names separated by a comma correspond to the 47 duplicates (see Materials and Methods).

    • ↵ b NT, nontypeable; two numbers separated by a comma mean that the strain reacted with both antisera.

    • ↵ c K. p., K. pneumoniae.

    • ↵ d The K serotypes of the clinical strains were determined in this study after cps PCR-RFLP analysis.

    • ↵ e ref., reference; CSF, cerebrospinal fluid; asp, aspirate; env., environment.

    • ↵ f Where source 1 is a reference strain, source 2 is the original strain name in the reference collection of the WHO reference center for E. coli and Klebsiella.

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Molecular Serotyping of Klebsiella Species Isolates by Restriction of the Amplified Capsular Antigen Gene Cluster
Sylvain Brisse, Sylvie Issenhuth-Jeanjean, Patrick A. D. Grimont
Journal of Clinical Microbiology Aug 2004, 42 (8) 3388-3398; DOI: 10.1128/JCM.42.8.3388-3398.2004

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Molecular Serotyping of Klebsiella Species Isolates by Restriction of the Amplified Capsular Antigen Gene Cluster
Sylvain Brisse, Sylvie Issenhuth-Jeanjean, Patrick A. D. Grimont
Journal of Clinical Microbiology Aug 2004, 42 (8) 3388-3398; DOI: 10.1128/JCM.42.8.3388-3398.2004
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    • ABSTRACT
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KEYWORDS

Genes, Bacterial
Klebsiella

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