Detection of Ureaplasma urealyticum in endotracheal tube aspirates from neonates by PCR

Abstract

A PCR-based test was optimized for the detection of Ureaplasma urealyticum from neonatal respiratory specimens, with primers directed against the multiple-banded antigen gene (L. J. Teng, X. Zheng, J. I. Glass, H. Watson, J. Tsai, and G. H. Cassell, J. Clin. Microbiol. 32:1464-1469, 1994). Endotracheal tube aspirates (225) from 103 low-birth-weight neonates (<1,250 g) were taken, when possible, at days 0, 4, and 14 after birth and examined by culture and by PCR. Of 77 specimens positive by either method, 73 were detected by PCR and 60 were detected by culture. Overall, 36% of the neonates were positive for U. urealyticum by either method. Of 16 patients with PCR-positive-culture-negative results, 13 had positive cultures at another sampling point, and one additional patient had a twin with positive cultures. Of 11 patients with day 0 specimens positive by PCR alone, 9 subsequently became culture positive, demonstrating the utility of this test in early detection. Multiple serovars were present in over 50% of positive specimens, with serovars 3 and 14 in combination being most prevalent. The amplicon size generated from the specimen by PCR correctly predicted the biovars isolated in over 85% of positive specimens. Thus, this PCR test was valuable in allowing early detection of U. urealyticum in neonatal respiratory specimens, as well as in providing biovar information.

FIG. 1

(A) Detection of U. urealyticum by two methodologies with all results over 14 days being shown. The total numbers of patients with ETTas positive for U. urealyticum by culture and by PCR at the three sampling points after birth are shown. The number of patients sampled is shown below each sampling point. (B) Detection of U. urealyticum by two methodologies, with only results for patients sampled at all three time points being shown. There were 45 patients who had samples taken at all three time points. The comparative results for these patients are shown.