Current and New Approaches in GMO Detection: Challenges and Solutions

Table 3

Representative examples illustrating multiplex qPCR TaqMan methods targeting GMO. Those validated at the EU level are indicated by an asterisk.

Multiplexing	Methods	Targets	References
Duplex	Element-specific	p35S and tNOS	[38]
Duplex	Element-specific	bar and pat	[41]
Duplex	Plant-specific	TLC	[42]
	Other	IPC
Duplex	Taxon-specific	ADH	[43]
	Event-specific	Bt11
Duplex	Taxon-specific	ADH	[43]
	Event-specific	Bt176
Duplex	Taxon-specific	ADH	[43]
	Event-specific	MON810
Duplex	Taxon-specific	ADH	[43]
	Event-specific	T25
Triplex	Element-specific	p35S, tNOS, and CTP2/CP4-EPSPS	[41]
Triplex	Taxon-specific	LEC and Zein	[42]
	Other	IPC
Triplex	Taxon-specific	Pro and PC	[42]
	Other	IPC
Triplex	Taxon-specific	ACC and FRUp	[42]
	Other	IPC
Triplex	Taxon-specific	SAD1 and FRUt	[42]
	Other	IPC
Triplex	Element-specific	p35S and pFMV	[42]
	Other	IPC
Triplex	Element-specific	tE9 and tNOS	[42]
	Other	IPC
Triplex	Element-specific	bar and CP4-EPSPS	[42]
	Other	IPC
Triplex	Element-specific	hpt and pat	[42]
	Other	IPC
Triplex	Element-specific	nptII and Cry1Ab/Ac	[42]
	Other	IPC
Triplex	Construct-specific	CBH351 and Bt176	[42]
	Other	IPC
Triplex	Construct-specific	MON810 and T25	[42]
	Other	IPC
Triplex	Construct-specific	Bt11 and MON863	[42]
	Other	IPC
Triplex	Construct-specific	NK603 and GA21	[42]
	Other	IPC
Triplex	Construct-specific	TC1507 and DAS-59122-7	[42]
	Other	IPC
Triplex	Construct-specific	MIR604 and MON88017	[42]
	Other	IPC
Triplex	Construct-specific	98140 and MON89034	[42]
	Other	IPC
Triplex	Construct-specific	3272 and MIR162	[42]
	Other	IPC
Triplex	Construct-specific	A2704-12 and GTS40-3-2	[42]
	Other	IPC
Triplex	Construct-specific	DP-305423-1 and DP-356043-5	[42]
	Other	IPC
Triplex	Construct-specific	MON87701 and MON89788	[42]
	Other	IPC
Triplex	Element-specific	AHAS	[42]
	Construct-specific	FG72
	Other	IPC	[42]
Triplex	Construct-specific	Bt63 and A5547-127	[42]
	Other	IPC
Triplex	Element-specific	Xa21
	Construct-specific	KMD1
	Other	IPC
Triplex	Taxon-specific	Zein	[44]
	Construct-specific	MON810 and GA21
Triplex	Taxon-specific	ADH	[44]
	Construct-specific	MON810 and GA21
Triplex	Element-specific	p35s, tNOS, and t35S	[45]
Triplex	Element-specific	tE9, pRbcS4, and tORF23	[45]
Triplex	Element-specific	tpinII and tAHASL	[45]
	Event-specific	DP-305423-1
Tetraplex	Element-specific	pFMV, bar, pat, and CTP2/CP4-EPSPS	[46]
Tetraplex	Element-specific	p35S, tNOS, pFMV, and bar	[47]
Pentaplex	Taxon-specific	HMG and LEC
	Element-specific	p35S and tNOS	[46]
	Virus-specific	CaMV
Pentaplex	Element-specific	p35S, tNOS, bar, pat, and CTP2/CP4-EPSPS	[41]
Pentaplex	Taxon-specific	LEC	[48]
	Event-specific	MON87769, MON87708, MON87705, and FG72
Hexaplex	Element-specific	p35S, tNOS, and pFMV
	Construct-specific	SAMS and LY	[49]
	Other	IPC

ACC (acetyl-CoA-carboxylase gene from colza); ADH (alcohol dehydrogenase I gene from maize); AHAS (AHAS fragment unique recombination from BPS-CV-127); bar (phosphinothricin-N-acetyltransferases gene from Streptomyces hygroscopicus); CaMV (ORFIII from CaMV); CP4-EPSPS (5-enolpyruvylshikimate-3-phosphate synthase gene from Agrobacterium tumefaciens strain); Cry (gene encoding the Bacillus thuringiensis δ-endotoxin); CTP2/CP4-EPSP (junction region between the chloroplast transit peptide 2 (CTP2) sequence from the Arabidopsis thaliana epsps gene and the CP4 epsps gene from Agrobacterium tumefaciens (CP4-EPSPS)); FRUp (β-fructosidase gene from potato); FRUt (β-fructosidase gene from tomato); HMG (major high-mobility group protein gene from maize); hpt (hygromycin phosphotransferase gene); IPC (internal positive control); LEC (lectin gene from soybean); LS28 (choline kinase); LY (transition from Zea mays chloroplast transit peptide sequence for dihydrodipicolinate synthase to Corynebacterium glutamicum dihydrodipicolinate synthase (cordapA) gene encoding for a lysine-insensitive dihydrodipicolinate synthase enzyme); nptII (neomycin phosphotransferase II gene); p35S (promoter of the 35 S cauliflower mosaic virus); pat (phosphinothricin-N-acetyltransferases gene from Streptomyces viridochromogenes); PC (phosphoenolpyruvate carboxylase gene from wheat); pFMV (promoter of the figwort mosaic virus); pRbcS4 (ribulose 1,5-bisphosphate carboxylase small subunit promoter from A. thaliana); Pro (prolamin gene from rice); SAD1 (stearoyl-acyl carrier protein desaturase gene from cotton); SAMS (transition from S-adenosyl-L-methionine synthetase (SAMS) promoter to Glycine max acetolactate synthase (gm-hra) gene); t35S (terminator of the cauliflower mosaic virus); tAHASL (acetohydroxy acid synthase large subunit terminator from A. thaliana); tE9 (ribulose-1,5-bisphosphate carboxylase terminator E9 from Pisum sativum); TLC (tRNA-Leu chloroplastic gene); tNOS (terminator of the nopaline synthase gene); tORF23 (open reading frame 23 terminator from A. tumefaciens); tpinII (inhibitor II terminator from potato); Zein (Zein gene from maize), Xa21 (Xa21 gene from Oryza longistaminata).