TABLE 3.

Procedures used for detection of C. pneumoniae in 2002

Lab no.	NA extraction method	NAAT	Detection method	Target	Monitoring inhibition	Monitoring of the procedurea		dUTP- UDG	Sample volume (μl)
						Neg. contr.	Pos. contr.		Extraction	Amplification
1	NucliSens Basic kit	In-house NASBA	ECL	16S rRNA	Y	Prep + Amp	Prep + Amp	N	100	5
1	NucliSens Basic kit	In-house NASBA	Real time	16S rRNA	Y	Prep + Amp	Prep + Amp	N	100	5
2	Roche Amplicor sputum	In-house PCR	Real time	PstI fragment	Y	None	None	Y	NS	5
3	QIAamp DNA	In-house PCR	Real time	16S rRNA gene	Y	Prep + Amp	Prep + Amp	N	200	10
4	Roche High Pure PCR Template Preparation kit	In-house PCR	Real time	PmP4 gene	Y	Prep + Amp	Amp	Y	200	10
5	QIAamp DNA	In-house PCR	Real time	MOMP gene	N	Prep + Amp	Prep + Amp	Y	NS	NS
6	QIAamp DNA blood	In-house PCR	Agarose gel	PstI fragment	Y	Prep + Amp	None	N	120	6
8	Roche High Pure PCR Template Preparation kit	In-house PCR	Real time	MOMP gene	N	Prep + Amp	Prep + Amp	Y	200	5
9	QIAamp DNA blood	In-house PCR	Real time	Cytadhesin gene	N	Amp	Amp	Y	200	5
10	In-house	In-house PCR	Real time	16S rRNA gene	N	Amp	Amp	Y	200	2
11	QIAamp DNA blood	In-house PCR	Real time	16S rRNA gene	N	Prep + Amp	Prep + Amp	Y	200	200
12	Roche High Pure PCR Template Preparation kit	In-house nested PCR	Agarose gel	MOMP gene	N	Prep + Amp	Prep + Amp	N	200	200
13	QIAamp DNA tissue	In-house PCR	Real time	PstI fragment	N	Amp	Amp	Y	200	5
14	Boiling	In-house PCR	Real time	PstI fragment	N	NS	Amp	Y	100	5
15	NucliSens Basic kit	In-house NASBA	Real time	16S rRNA	N	None	None	N	100	5
15	NucliSens Basic kit	In-house NASBA	Real time multiplex	16S rRNA	N	None	None	N	100	5

^aAmp, amplification; ECL, electrochemiluminescent detection; dUTP-UDG, deoxyuridine triphosphate-uracyl-N-glycosylase (dUTP replaces dTTP in PCR reactions, and the enzyme excises uracyl from any contaminating PCR product to prevent false positives); N, no; NAAT, nucleic acid amplification test; NASBA, nucleic acid sequence-based amplification; prep, preparation; Y, yes; NA, nucleic acid; neg., negative; pos., positive; contr., control; NS, not specified.