Article Figures & Data
Figures
- 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.
- 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
- Table 1.
Sexes and ages of HIV-positive children and HIV-negative controls
Parameter HIV-positive children HIV-negative controls No. of children 15 15 No. of males:no. of females 8:7 13:2 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 × g 61.1 2,600 × g 33.9 Growth medium Broth 51.4 Agar 48.6 Type of growth medium SP4 37.0 HF 19.2 Friis 19.2 PH 24.6 Other methods Blind passagea 26.4 Plate washb 8.3 Colony picksc 65.3 ↵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 classification No. of patients Mean (range) age (yr) Median CD4 count (no. of cells/mm3) N 1 11.5 744 A 3 6.0 (2.5–6.0) 300 B 2 2.7 (0.6–4.10) 654 C 9 5.0 (1.4–13.3) 280 - Table 4.
Preliminary identification of mycoplasmas isolated from pediatric patients
Mycoplasma species Biochemical test result PCR result Glucose utiliza-tion Arginine hydro-lysis Urease hydro-lysis Size (bp) of 16S-23S rRNA product HindIIIb VspIb 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-fication Median CD4 count (no. of cells/mm3) Mycoplasma species isolated 2.3 C2 696 M. hominis, M. penetrans 6.0 A3 90 M. hominis, U. urealyticum 9.8 C2 330 M. hominis, U. urealyticum 9.8 C2 280 M. hominis,M. penetrans 11.0 C3 30 M. hominis,M. penetrans - Table 6.
Distribution of patients according to degree of immunologic suppression, age, and CDC classification
Degree of immunologic suppression No. of patients with the following CDC classification: No. of patients of the following age (median CD4 count [no. of cells/mm3]): N A B C <12 mo 1–5 yr >6 yr None 1 1 1 2 (1,425) 1 Moderate 1 2 4 1 (859) 2 (572) 4 Severe 1 4 2 (98) 3 - Table 7.
Mycoplasma species isolated from HIV-negative controls and HIV-positive children according to CDC classification
CDC classification No. of patients No. (%) of each species isolated M. hominis U. urealyticum M. genitalium M. pirum M. fermentans M. penetrans Control 15 3 (20) 3 (20) N 1 1 (100) A 3 2 (67) 2 (67) B 2 1 (50) C 9 4 (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 patients Median CD4 count (no. of cells/mm3) No. (%) of each species isolated M. hominis U. urealyticum M. genitalium M. pirum M. fermentans M. penetrans <1 1 859 1–5 6 572 1 (17) 1 (17) 1 (17) 2 (33) >6 8 290 6 (75) 3 (38) 1 (13) 4 (50)
