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Bacteriology

Mycoplasma penetrans and Other Mycoplasmas in Urine of Human Immunodeficiency Virus-Positive Children

Althaf I. Hussain, William Lane M. Robson, Robin Kelley, Tanya Reid, J. David Gangemi
Althaf I. Hussain
Department of Microbiology and Molecular Medicine, Clemson University, Clemson, South Carolina;
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William Lane M. Robson
Department of Pediatric Nephrology and
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Robin Kelley
Department of Pediatric Infectious Disease, The Children’s Hospital, Greenville Hospital System, and
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Tanya Reid
Children’s Immunology Center,University of South Carolina School of Medicine, Columbia, South Carolina
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J. David Gangemi
Greenville Hospital System/Clemson University Biomedical Cooperative, Greenville, South Carolina; and
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DOI: 10.1128/JCM.37.5.1518-1523.1999
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  • Fig. 1.
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    Fig. 1.

    Agarose gel electrophoresis of the second-step PCR products. 16S-23S rRNA PCR of intergenic spacer regions ofMycoplasma species isolated from urine sediments. Ten microliters of the second-step PCR products was subjected to electrophoresis in a 2% agarose gel. Lane 1, 50- to 2,000-bp DNA marker; lane 2, M. pirum; lane 3, M. fermentans; lane 4, M. penetrans; lane 5, M. hominis; lane 6,M. salivarium ATCC 23064; lane 7, negative control.

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    Fig. 2.

    Restriction enzyme digestion pattern of the second-step PCR products. Five microliters of the second-step products was digested with 1 U of HindIII or VspI, electrophoresed in a 2% agarose gel, and stained with ethidium bromide. Lanes 1 and 8, 2,000- to 50-bp DNA marker; lanes 2, M. penetrans; lanes 3,M. fermentans; lanes 4, M. genitalium; lanes 5,M. pirum; lanes 6, M. hominis; lanes 7, M. salivarium ATCC 23064, used as a positive control. (A)HindIII-cleaved PCR products; (B)VspI-cleaved PCR products.

Tables

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  • Table 1.

    Sexes and ages of HIV-positive children and HIV-negative controls

    ParameterHIV-positive childrenHIV-negative controls
    No. of children1515
    No. of males:no. of females8:713:2
    Median (range) age (yr)5.6 (0.6–13.3)9 (4.0–16.0)
  • Table 2.

    Isolation of mycoplasmas from urine sediments

    Characteristic% Isolation
    Centrifugation speed
      500 × g61.1
      2,600 × g33.9
    Growth medium
     Broth51.4
     Agar48.6
    Type of growth medium
     SP437.0
     HF19.2
     Friis19.2
     PH24.6
    Other methods
     Blind passagea26.4
     Plate washb8.3
     Colony picksc65.3
    • ↵a At least two blind passages were done with samples which showed no signs of growth before a sample was considered negative for mycoplasmas.

    • ↵b When organisms could not be grown from agar blocks, liquid medium was transferred to those plates, and after 24 h of incubation, the liquid medium was aspirated from the agar surface and transferred into broth cultures to obtain stocks.

    • ↵c Agar surfaces showing a colony morphology with a typical fried-egg appearance were cut and transferred into broth cultures.

  • Table 3.

    Number of patients, mean age of patients, and median CD4 count according to CDC classification

    CDC classificationNo. of patientsMean (range) age (yr)Median CD4 count (no. of cells/mm3)
    N111.5744
    A36.0 (2.5–6.0)300
    B22.7 (0.6–4.10)654
    C95.0 (1.4–13.3)280
  • Table 4.

    Preliminary identification of mycoplasmas isolated from pediatric patients

    Mycoplasma speciesBiochemical test resultPCR result
    Glucose utiliza-tionArginine hydro-lysisUrease hydro-lysisSize (bp) of 16S-23S rRNA productHindIIIbVspIb
    M. penetrans++−295−−
    M. fermentans++−364++
    M. pirum++−323++
    M. genitalium+−−253−+
    M. hominis−+−236−+
    • a Second PCR products.

    • ↵b Digestion with restriction enzymesHindIII and VspI.

  • Table 5.

    Age, CDC classification, and median CD4 count for patients from whom more than one Mycoplasma species was isolated

    Age (yr)CDC classi-ficationMedian CD4 count (no. of cells/mm3)Mycoplasma species isolated
    2.3C2696M. hominis, M. penetrans
    6.0A390M. hominis, U. urealyticum
    9.8C2330M. hominis, U. urealyticum
    9.8C2280M. hominis,M. penetrans
    11.0C330M. hominis,M. penetrans
  • Table 6.

    Distribution of patients according to degree of immunologic suppression, age, and CDC classification

    Degree of immunologic suppressionNo. of patients with the following CDC classification:No. of patients of the following age (median CD4 count [no. of cells/mm3]):
    NABC<12 mo1–5 yr>6 yr
    None1112 (1,425)1
    Moderate1241 (859)2 (572)4
    Severe142 (98)3
  • Table 7.

    Mycoplasma species isolated from HIV-negative controls and HIV-positive children according to CDC classification

    CDC classificationNo. of patientsNo. (%) of each species isolated
    M. hominisU. urealyticumM. genitaliumM. pirumM. fermentansM. penetrans
    Control153 (20)3 (20)
    N11 (100)
    A32 (67)2 (67)
    B21 (50)
    C94 (44)1 (11)1 (11)1 (11)1 (11)5 (55.5)
  • Table 8.

    Mycoplasma species isolated from HIV-positive children according to age

    Age (yr)No. of patientsMedian CD4 count (no. of cells/mm3)No. (%) of each species isolated
    M. hominisU. urealyticumM. genitaliumM. pirumM. fermentansM. penetrans
    <11859
    1–565721 (17)1 (17)1 (17)2 (33)
    >682906 (75)3 (38)1 (13)4 (50)
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Mycoplasma penetrans and Other Mycoplasmas in Urine of Human Immunodeficiency Virus-Positive Children
Althaf I. Hussain, William Lane M. Robson, Robin Kelley, Tanya Reid, J. David Gangemi
Journal of Clinical Microbiology May 1999, 37 (5) 1518-1523; DOI: 10.1128/JCM.37.5.1518-1523.1999

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Mycoplasma penetrans and Other Mycoplasmas in Urine of Human Immunodeficiency Virus-Positive Children
Althaf I. Hussain, William Lane M. Robson, Robin Kelley, Tanya Reid, J. David Gangemi
Journal of Clinical Microbiology May 1999, 37 (5) 1518-1523; DOI: 10.1128/JCM.37.5.1518-1523.1999
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KEYWORDS

Acquired Immunodeficiency Syndrome
bacteriuria
HIV Seropositivity
mycoplasma

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