Circulating and stool nucleic acid analysis for colorectal cancer diagnosis

Table 2 Summary of studies that have tested DNA integrity and genetic alteration markers in stool samples

Study	Biomarker(s)	Methods	Assay for Long DNA	Case analyzed	Sensitivity	Specificity
Ahlquist et al[88], 2000	Stool DNA integrity	L-DNA (4 sites)	Hybrid	CRC: 22	91%	93%
	KRAS, TP53, APC	Mutation analysis	Capture	A ≥ 1 cm: 11	82%	NA
	BAT 26	MSI	PCR	HD: 28
Tagore et al[89], 2003	Stool DNA integrity	L-DNA (6 sites)	Hybrid	CRC: 52	64%	96%
	KRAS, TP53, APC	Mutation analysis	Capture	AA: 28
	BAT 26	MSI	PCR	HD: 212	57%	NA
Calistri et al[90], 2003	Stool DNA integrity	L-DNA (8 sites)	PCR	CRC: 56	62%	97%
	KRAS, TP53, APC	Mutation analysis		HD: 38
	D2S123, D5S346, D17S250
	BAT 25, BAT 26	MSI
Boyton et al[91], 2003	Stool DNA integrity	L-DNA (6 sites)	Hybrid	CRC: 27	56%	97%
			Capture	HD: 77
			PCR
Calistri et al[92], 2004	Stool DNA integrity	FL-DNA (8 sites)	Capillary	CRC: 85	76%	93%
			Electrophoresis	HD: 59
Whitney et al[93], 2004	Stool DNA integrity	L-DNA (4 sites)	Magnetic	CRC: 86	70%	96%
	KRAS, TP53, APC	Mutation analysis	Bead-Based	HD: 100
	BAT 26	MSI	Sequence-Specific Purification
			Capture
Imperiale et al[94], 2004	Stool DNA integrity	L-DNA (4 sites)	Hybrid Capture	CRC: 31	52%	95%
	KRAS, TP53, APC	Mutation analysis	PCR	AA: 407	15%	NA
	BAT 26	MSI		P: 648	8%	NA
				HD: 1423
Kutzner et al[95], 2005	Stool DNA integrity	L-DNA (4 sites)	Hybrid	CRC: 57	65%	91%
	APC	Mutation analysis	Capture	HD: 44
	BAT 26	MSI	PCR
Zou et al[96], 2006	Stool DNA integrity	L-DNA: Alu-assay (2 sites)	RT PCR	CRC: 18	44%	100%
				HD: 20
Itzkowitz et al[97], 2007	Stool DNA integrity	L-DNA (4 sites Locus D, Locus Y)	RT PCR	CRC: 40	88%	82%
	Vimentin	Methylation analysis		HD: 122
Abbaszadegan et al[98], 2007	Stool DNA integrity	L-DNA: (1476 bp fragments)	PCR	CRC: 25	64%	95%
	p16	Methylation analysis		HD: 20
	BAT 26	MSI
Ahlquist et al[99], 2008	Stool DNA integrity	L-DNA (4 sites)	PCR	CRC: 12	25%	96%
	KRAS, APC, TP53	Mutations Analysis		A ≥ 1 cm: 135	17%	NA
	BAT 26	MSI		A < 1 cm: 469
	Stool DNA test 1			P: 341	4%	NA
				HD: 1473	5%	NA
Itzkowitz et al[47], 2008	Stool DNA integrity	L-DNA ( 4 sites, Locus D, Locus Y)	RT PCR	CRC: 82	83%	82%
	Vimentin	Methylation Analysis		HD: 363
Calistri et al[100], 2009	Stool DNA integrity	FL-DNA (8 sites)	Capillary	CRC: 100	79%	89%
			Electrophoresis	HD: 100
Calistri et al[101], 2010	Stool DNA integrity	FL-DNA (8 sites)	Capillary	CRC: 26	Cancer risk prediction with markers combination
	iFOBT	iFOBT	Electrophoresis	A HR: 264
				A LR: 54
				HD: 216
Kalimutho et al[102], 2011	Stool DNA integrity	L-DNA (4 sites)	QdHPLC	CRC: 28	86%	81%
	Calprotectin	ELISA		A: 69	17%	NA
				HD: 95	72%	75%
Ahlquist et al[103], 2012	Stool DNA integrity	L-DNA	QuARTS	CRC: 252	85%	89%
	KRAS	Mutations Analysis		A ≥ 1 cm: 133	63%	NA
	NDRG4, BMP3, vimentin,	Methylation Analysis		A < 1 cm: 94	54%	NA
	TFP12/α-actine			HD: 293

L-DNA: Long-DNA; FL-DNA: Fluorescence long DNA; CRC: Colorectal cancer; A: Adenoma; AA: Advanced Adenoma; A HR: Adenoma high risk; A LR: Adenoma low risk; HD: Healthy donors; HP: Hyperplasic polyps; NA: Not available; RT-PCR: Real time polymerase chain reaction; QdHPLC: Quantitative denaturing high performance liquid chromatografy; QuARTS: Quantitative allele specific real-time target and signal amplification; MSI: Microsatellite instability.