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Bacteriology

Polymorphism in the Pertussis Toxin Promoter Region Affecting the DNA-Based Diagnosis of BordetellaInfection

Malin Nygren, Elisabet Reizenstein, Mostafa Ronaghi, Joakim Lundeberg
Malin Nygren
Department of Biotechnology, KTH, Royal Institute of Technology, and
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Elisabet Reizenstein
Department of Bacteriology, Swedish Institute for Infectious Disease Control, Stockholm, Sweden
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Mostafa Ronaghi
Department of Biotechnology, KTH, Royal Institute of Technology, and
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Joakim Lundeberg
Department of Biotechnology, KTH, Royal Institute of Technology, and
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DOI: 10.1128/JCM.38.1.55-60.2000
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    Fig. 1.

    Results of agarose gel electrophoresis demonstrating the two different fragment sizes obtained with PCR primers STMA17 and STMA18. Lane 1, amplified product of B. pertussis patient isolate KP92\2\92; lane 2, B. pertussis ATCC 9797; lane 3,B. parapertussis ATCC 15311; lane 4, B. bronchiseptica ATCC 786; lane 5, B. bronchiseptica19395; lane M, a 50-bp ladder.

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    Fig. 2.

    The five different sequences encompassing the PT promoter region (the 239 or 249 nucleotides upstream of the start codon of the PT subunit 1 gene amplified with flanking primers) found among the 33 ATCC reference strains and patient isolates sequenced. The three restriction sites used for species identification are marked with boxes. Restriction enzyme recognition sequences are underlined. The nucleotide positions in sequence b (B. pertussis ATCC 9797 and ATCC 9340) that differ from those in sequence a (B. pertussis patient isolates) are marked in boldface. The nucleotide positions in sequence e (B. bronchiseptica ATCC 19395) that differ from those in sequence d (B. bronchiseptica ATCC 786 and AB 1254) are also marked in boldface. Sequence c is the sequence common to all of the B. parapertussis strains.

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    Fig. 3.

    The four different restriction patterns resulting from the five different sequence types in Fig. 2 obtained with restriction enzymes TaqI, BglI, and HaeII.

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    Fig. 4.

    Results from colorimetric restriction enzyme analysis ofBordetella strains representing the five different PT promoter sequence types in Fig. 2 to confirm the four restriction patterns in Fig. 3. The negative control corresponds to a PCR-negative sample that was immobilized onto magnetic beads and mixed with fusion protein and substrate (see text for details).

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  • Table 1.

    Strains used in the study

    Strain or patient isolateSequence typeRestriction patternSourceYear of isolation
    B. pertussis
     KP92\2\92aIPVTa92-951992
     KP92\9\92aIPVT 92-951992
     KP92\12\92aIPVT 92-951992
     KP92\42\92aIPVT 92-951992
     KP92\47\92aIPVT 92-951992
     KP92\357\93aIPVT 92-951993
     KP92\757\94aIPVT 92-951994
     KP92\1145\94aIPVT 92-951994
     KP92\1162\94aIPVT 92-951994
     R613257 SBL 77-09-29aIRoutine clinical sample1977
     RA776 SBL 75-10-02aIRoutine clinical sample1975
     ATCC 9340bIATCC
     ATCC 9797bIATCC
    B. parapertussis
     KP92\11\92cIIPVT 92-951992
     KP92\41\92cIIPVT 92-951992
     KP92\83\93cIIPVT 92-951993
     KP92\106\93cIIPVT 92-951993
     KP86\148\86cIIPVT 86-871986
     KP92\214\93cIIPVT 92-951993
     KP92\218\93cIIPVT 92-951993
     KP86\250\86cIIPVT 86-871986
     KP92\283\93cIIPVT 92-951993
     KP92\308\93cIIPVT 92-951993
     KP92\373\93cIIPVT 92-951993
     KP92\377\93cIIPVT 92-951993
     KP92\437\93cIIPVT 92-951993
     KP92\756\94cIIPVT 92-951994
     KP92\1180\94cIIPVT 92-951994
     ATCC 15237cIIATCC
     ATCC 15311cIIATCC
    B. bronchiseptica
     ATCC 786dIIIATCC
     AB 1254dIIISSb
     ATCC 19395eIVATCC
    • ↵a PVT, pertussis vaccine trials, 1986 to 1987 (PVT 86-87) (1) and 1992 to 1995 (PVT 92-95) (13).

    • ↵b SS, Statens Seruminstitut, Copenhagen, Denmark.

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Polymorphism in the Pertussis Toxin Promoter Region Affecting the DNA-Based Diagnosis of BordetellaInfection
Malin Nygren, Elisabet Reizenstein, Mostafa Ronaghi, Joakim Lundeberg
Journal of Clinical Microbiology Jan 2000, 38 (1) 55-60; DOI: 10.1128/JCM.38.1.55-60.2000

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Polymorphism in the Pertussis Toxin Promoter Region Affecting the DNA-Based Diagnosis of BordetellaInfection
Malin Nygren, Elisabet Reizenstein, Mostafa Ronaghi, Joakim Lundeberg
Journal of Clinical Microbiology Jan 2000, 38 (1) 55-60; DOI: 10.1128/JCM.38.1.55-60.2000
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KEYWORDS

Bordetella
Bordetella Infections
Pertussis Toxin
Polymorphism, Genetic
Promoter Regions, Genetic
Virulence Factors, Bordetella

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