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Mycobacteriology and Aerobic Actinomycetes

Comparison of Roche Cobas Amplicor Mycobacterium tuberculosis Assay with In-House PCR and Culture for Detection of M. tuberculosis

Bodo R. Eing, Andrea Becker, Arthur Sohns, Ronald Ringelmann
Bodo R. Eing
Institut für Medizinische Mikrobiologie und Immunologie, Städtisches Klinikum Karlsruhe, 76133 Karlsruhe, Germany
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Andrea Becker
Institut für Medizinische Mikrobiologie und Immunologie, Städtisches Klinikum Karlsruhe, 76133 Karlsruhe, Germany
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Arthur Sohns
Institut für Medizinische Mikrobiologie und Immunologie, Städtisches Klinikum Karlsruhe, 76133 Karlsruhe, Germany
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Ronald Ringelmann
Institut für Medizinische Mikrobiologie und Immunologie, Städtisches Klinikum Karlsruhe, 76133 Karlsruhe, Germany
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DOI: 10.1128/JCM.36.7.2023-2029.1998
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    Fig. 1.

    Results of conventional testing of noninhibitory specimens.

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  • Table 1.

    Comparison of CA and in-house PCR with culture

    SpecimensAmplification systemNo. of culture-positive specimensa that were PCR:No. of culture-negative specimensb that were PCR:Sensitivity (%)Specificity (%)NPV (%)PPV (%)
    PositiveNegativePositiveNegative
    All (n= 1,527)CA5015211,44176.9298.5698.9770.42
    In house596351,42790.7697.6099.5862.72
    Smear positiveCA2711396.4275.0075.0096.42
     (n = 32)cIn house28031100.0025.00100.0090.32
    Smear negativedCA2314201,43862.1698.5699.0353.48
     (n = 1,495)In house316321,42683.7897.6099.5846.20
    • ↵a Includes only specimens growingM. tuberculosis.

    • ↵b Includes 10 culture-positive specimens growing MOTT.

    • ↵c Includes specimens from TB patients only.

    • ↵d Includes six unspecific positive smears and 10 smear-positive specimens from patients infected with MOTT.

  • Table 2.

    Adjustment of discrepant results to the adapted gold standard

    TestNo. of true-positive specimens for which:No. of specimens for which positive PCR was not confirmableb
    Positive PCR was confirmed by other culture-positive samples from same patientPositive PCR was confirmed by other PCRNegative PCR was confirmed by both PCRsa
    CA1524
    In-house PCR3122
    Acid-fast staining1
    • ↵a Smear-negative, culture-negative specimen from a patient with culture-proven TB.

    • ↵b False-positive PCR.

  • Table 3.

    Comparison of CA and in-house PCR to the adapted gold standard

    SpecimensAmplification systemNo. of adapted gold standard-positive specimensa that were PCR:No. of adapted gold standard-negative specimensb that were PCR:Sensitivity (%)Specificity (%)NPV (%)PPV (%)
    PositiveNegativePositiveNegative
    All (n= 1,527)CA673441,42266.3399.7197.6694.36
    In house92921,42491.0899.8599.3797.87
    Smear positiveCA2840087.5099.37
     (n = 32)cIn house3110096.8799.37
    Smear negativeCA393041,42256.5299.7197.9390.69
     (n = 1,495)dIn house61821,42488.4099.8599.4496.82
    • ↵a Specimens from patients with TB (for a definition, see Materials and Methods).

    • ↵b Specimens from patients without any evidence of TB.

    • ↵c Includes only specimens from TB patients.

    • ↵d Includes six unspecific positive smears and 10 smear-positive specimens from patients infected with MOTT.

  • Table 4.

    Sensitivity of CA and in-house PCR compared to culture with different types of specimens

    Specimen typeNo. of all samples (sensitivity [%]) that were:No. of smear-negative samples (sensitivity [%]) that were:
    Culture positiveCA positiveIn-house PCR positiveCulture positiveCA positiveIn-house PCR positive
    Sputum or BALa2317 (73.91)19 (82.60)115 (45.45)7 (63.63)
    Gastric aspirate3427 (79.41)32 (94.11)2014 (70.00)18 (90.00)
    Urine44 (100.00)4 (100.00)44 (100.00)4 (100.00)
    Other42 (50.00)4 (100.00)20 (0)2 (100.00)
    • ↵a BAL, bronchoalveolar lavage.

  • Table 5.

    Inhibition rates of CA with different specimen types

    Specimen typeaTotal no. of specimensNo. of noninhibitory specimensNo. of inhibitory specimensInhibition rate (%)
    Pus474524.25
    Swab323113.12
    Sputum or BALa8217338810.71
    Gastric aspirate3543094512.71
    Urine15715342.54
    Biopsy868333.48
    Pleural fluid12211397.37
    CSFb292900.00
    Other333126.06
     Total1,6811,5271549.16
    • ↵a BAL, bronchoalveolar lavage.

    • ↵b CSF, cerebrospinal fluid.

  • Table 6.

    Role of PCR for the primary diagnosis of TB and specification of positive smear results

    Test(s) performed for primary diagnosisNo. of specimensComment
    Microscopy plus CA or in-house PCR4Smear-positive MOTT infections
    4Unspecific smear results
    17Smear-positive TB cases
    CA or in-house PCR7Smear-negative TB cases (including 2 culture-negative cases)
    In-house PCR only4Smear-negative TB cases (including 3 missed by CA)
    In-house PCR2False-positive PCR results
    CA4False-positive PCR results
     Total42Cases with a positive result by any of the investigated methods
  • Table 7.

    Assessment of the minimum number of specimens necessary for CA for smear-negative TB patients

    PatientNo. of specimensaNo. of true-positive specimensbRate of true-positive specimens (%)No. of CA-positive specimensSequential no. of first CA-positive specimencLocalization of infection
    554410041Urogenital organs
    5435510021Lungs
    2665510021Lungs
    1761110011Lungs
    2221110011Lymph node
    5053310022Lungs
    2871011017Lungs
    3569555.618Lungs
    354331000Lungs
    5366116.70Mediastinal lymph node
    5873133.30Pulmonary lymph node
    • ↵a Includes only specimens drawn from the site of infection before initiation of therapy. For details, see Materials and Methods.

    • ↵b See also the definition of the adapted gold standard in Materials and Methods.

    • ↵c As determined after ordering the specimens of each patient by the time of receipt.

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Comparison of Roche Cobas Amplicor Mycobacterium tuberculosis Assay with In-House PCR and Culture for Detection of M. tuberculosis
Bodo R. Eing, Andrea Becker, Arthur Sohns, Ronald Ringelmann
Journal of Clinical Microbiology Jul 1998, 36 (7) 2023-2029; DOI: 10.1128/JCM.36.7.2023-2029.1998

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Comparison of Roche Cobas Amplicor Mycobacterium tuberculosis Assay with In-House PCR and Culture for Detection of M. tuberculosis
Bodo R. Eing, Andrea Becker, Arthur Sohns, Ronald Ringelmann
Journal of Clinical Microbiology Jul 1998, 36 (7) 2023-2029; DOI: 10.1128/JCM.36.7.2023-2029.1998
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KEYWORDS

Mycobacterium tuberculosis
polymerase chain reaction
sputum
Tuberculosis, Pulmonary

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