Table 3.

Summary of recent mono polymerase chain reaction (PCR) assays for the detection of M. pneumoniae published since 2003 and previously validated assays used as comparators

Reference	Assay type	Detection format	Target gene (bp)	PCR assay used as a comparator for the new assay	Non-PCR comparator test	Specimens tested for the validation of sensitivity and/or specificity
2003 [71]	PCR	Molecular beacons	P1 gene (151)	[72, 73]	Serology	Various bacterial species, DNA dilutions, clinical specimens
2004 [70]	PCR	Agarose gel electrophoresis	16S rRNA gene (225)	ND	Culture, serology	M. pneumoniae dilution series
2004 [74]	Broad-range PCR	Microarray	parE gene (±300)	ND	ND	Various bacterial species, DNA dilutions, clinical specimens
2004 [60]	Nested PCR	Reverse line blot hybridization	16S-23S rRNA spacer (94)	ND	ND	21 Mollicute reference strains, 92 contaminated cell cultures, 80 Mollicute isolates, 14 M. pneumoniae-positive NPAs, 6 M. pneumoniae-negative specimens Pathogens targeted: M. arginini, M. fermentans, M. hyorhinis, M. orale, Acholeplasma laidlawii, M. pneumoniae, M. hominis, M. genitalium, Ureaplasma parvum, U. urealyticum
2005 [75]	PCR	Real-time	16S RNA gene (NS)	ND	Culture	Spiked sputa and BALs, clinical specimens with known M. pneumoniae status by culture
2005 [76]	LAMP	Turbidimeter	P1 gene (NS)	[77]	ND	Various bacterial species, DNA dilutions, clinical specimens
2006 [78]	PCR	Molecular beacon	16S rRNA gene (225)	ND	Culture, serology	Various bacterial species, bacterial dilution series, clinical specimens
2006 [79]	PCR	Real-time	P1 gene (141)	ND	Culture, serology	Various bacterial strains, dilutions of cloned DNA, clinical specimens with known M. pneumoniae status by culture and serology
2007 [80]	PCR	Scorpion probe	P1 gene (72)	[81, 82]	Serology	Various bacterial strains, dilutions of M. pneumoniae DNA, clinical specimens with unknown status
2007 [83]	PCR	Real-time	repMp1 in P1 (184)	P1 gene-based PCR (177 bp)	ND	Various bacterial species, dilutions of plasmids containing target sequence, clinical specimens with known M. pneumoniae status by PCR
2008 [51]	PCR	Real-time	CARDS toxingene (73) ATPase gene(68) ATPase gene(106)	ND	ND	Various bacterial species, bacterial dilution series, clinical specimens from an outbreak
2009 [59]	Broad-range PCR	Real-time	tuf gene (160)	VenorGeM-DI Mycoplasma detection kit, MycoSensor QPCR Assay kit	ND	32 Mollicute species, various other bacterial species, cell culture supernatants, clinical specimens (80sputa, 5 throat swabs)

A-test: passive agglutination-test; LAMP: loop-mediated isothermal amplification; ND: not done; MycoSensor QPCR Assay (Stratagene, LaJolla, CA, USA); VenorGeM-DI Mycoplasma detection kit (Minerva Biolabs GmbH, Berlin, Germany)