InMycoplasma hominis, two genes (alrandgoiB) have been found to be associated with the invasion of the amniotic cavity, and a single gene (goiC) to be associated with intra-amniotic infections and a high risk of preterm birth. The syntopic presence ofUreaplasmaspp. in the same patient has been shown to correlate with the absence ofgoiCinM. hominis. The aim of our study was to investigate the presence ofalr,goiB, andgoiCgenes in two groups ofM. hominisisolates collected from symptomatic and asymptomatic male and non-pregnant female patients attending an Outpatients Centre. Group A consisted of 26 isolates from patients with onlyM. hominisconfirmed; group B consisted of 24 isolates from patients withUreaplasmaspp. as the only co-infection. We extracted DNA from allM. hominisisolates and analysed the samples for the presence ofalr,goiB, andgoiCin a qPCR assay. Additionally, we determined their cytotoxicity against HeLa cells. We confirmed the presence of thealrgene in 85% of group A isolates and in 100% of group B isolates;goiBwas detected in 46% of the samples in both groups, whereasgoiCwas found in 73% of group A and 79% of group B isolates, respectively. It was shown that co-colonisation withUreaplasmaspp. in the same patient had no effect on the presence ofgoiCin the respectiveM. hominisisolate. We did not observe any cytotoxic effect of the investigated isolates on human cells, regardless of the presence or absence of the investigated genes.