Two duplex PCR assays were established for the detection of C. trachomatis (Ct), N. gonorrhoeae
(GC), M. hominis (Mh), and U. urealyticum (Uu). These assays were used on clinical specimens
obtained from women with Premature Rupture of Membrane or Post Partum Fever, from preterm
infants, as well as from women with uneventful pregnancies and their babies delivered vaginally
at term. The analytical sensitivity of the duplex PCR assays with internal controls incorporated is 7.0,
19.0, 5x103 and 7x102 genome copies per reaction for Ct, GC, Mh and Uu respectively. Specificity
was demonstrated by the amplification of only target DNA in the presence of other organisms.
Among 40 women with normal, at term, deliveries, there were 6 positives for Ct, 2 for GC and 1
for Uu. None of these women had signs of genital tract infection. The Mh/Uu PCR was positive in
11 of 40 PROM cases, with 7 women positive for Uu, 2 for Mh and 2 others for both organisms.
Of 40 blood cultures taken from post-partum maternal infections, 6 were positive for Ct and 1 for
Mh. Respiratory secretions from 30 premature neonates yielded 5 positives for Uu and one each
for Mh and Ct. In contrast, there was only 1 positive result (for Mh) in 30 mature neonates. With 1
exception, all mycoplasma and ureaplasma positives were confirmed by culture and the concordance
between paired tracheal aspirates and nasopharyngeal swabs from neonates was 96.7%. These results show the potential use of the duplex PCR assays for the diagnosis of maternal and neonatal disease caused by the four urogenital pathogens.