Abstract
Megasphaera micronuciformis is an anaerobic microbe isolated from humans. However, since the microbe is strictly anaerobic, its cultivation requires complicated facilities, making detection costly. For rapid, inexpensive detection and identification of M. micronuciformis in the clinical setting, a new technique is necessary. This study aimed to develop a species-specific PCR primer set for the detection of M. micronuciformis. A ribosomal DNA-specific PCR primer Mm2F was designed for M. micronuciformis. Analytical specificity data showed that the PCR primer set Mm2F/Mega-X produced amplicons from M. micronuciformis but not from the other species tested, including 4 Megasphaera species and representative related species. Of the 52 oral samples from Japanese subjects evaluated in our study, 71% were positive for M. micronuciformis, suggesting the likelihood that M. micronuciformis is widely distributed in the oral cavity of the Japanese population. Key words: Megasphaera micronuciformis, specific polymerase chain reaction (PCR), Mm2F, oral bacteria.
Highlights
Megasphaera micronuciformis is a Gram-negative, anaerobic microorganism that was first isolated from a liver abscess and pus sample in adult women (Marchandin et al, 2003)
This study aimed to develop a species-specific polymerase chain reaction (PCR) primer set for the detection of M. micronuciformis
A ribosomal DNA-specific PCR primer Mm2F was designed for M. micronuciformis
Summary
Megasphaera micronuciformis is a Gram-negative, anaerobic microorganism that was first isolated from a liver abscess and pus sample in adult women (Marchandin et al, 2003). It belongs to the phylum Firmicutes and genus Megasphaera, which includes Megasphaera cerevisiae, Megasphaera elsdenii, Megasphaera paucivorans and Megasphaera sueciensis (Haikara and Helander, 2006; Vos et al, 2009; Ohnishi et al, 2010). Since its culture is difficult (Kumar et al, 2005), the abovementioned characteristics must be investigated using culture-independent methodologies such as cloning, microarray and pyro sequencing (Riggio et al, 2008; Preza et al, 2009; Guss et al, 2010). Since an effective tool for M. micronuciformis detection is necessary, we aimed to develop a polymerase chain reaction (PCR) primer set for use in rapid M. micronuciformis detection
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