Detection of Myocardial Injury Markers Based on Double Gold Nanoparticles Probes and Protein Chip†

doi:10.7503/cjcu20150133

Abstract

Abstract:

A novel method based on gold nanoparticles(AuNPs) probes and protein chip was developed to detect myocardial injury markers. Two AuNPs probes were constructed: one was modified with detection antibody and DNA probe 1(detection probe), and the other was labeled with DNA probe 2 which was complementary to DNA probe 1(signal probe). When target antigen was present, a sandwich structure(capture antibody-target antigen-detection probe-signal probe) was formed through the interaction of the antibody-antigen and the hybridization of complemenatry DNAs. Then the chip was dyed by the gold deposition solution, and the signal was amplified. The results were observed with a microscope linked with an image analysis software. This system could simultaneously detect multiple markers in 40 min. The detection limit of cTnⅠ was 10 pg/mL, which was equivalent to ECLIA in clinical. For HFABP and MYO, the detection limit could be as low as 640 and 10 pg/mL, respectively, and the sensitivity was improved greatly when compared with ECLIA and ELISA.

Key words: Gold nanoparticles probe, Protein chip, Hybridization, Myocardial injury marker

CLC Number:

O657.39

TrendMD:

HUANG Lijun, ZHANG Lihua, MAO Hongju, WANG Ping, JIANG Youxu, LI Panpan, JIN Qinghui, ZHAO Jianlong. Detection of Myocardial Injury Markers Based on Double Gold Nanoparticles Probes and Protein Chip^†[J]. Chem. J. Chinese Universities, 2015, 36(9): 1687.

Figures/Tables 10

Scheme 1 Schematic of the detection of myocardial injury markers based on double gold nanoparticle probes and protein chip

Fig.1 TEM images of bare AuNPs(A) and AuNPs modified with detection antibodies and DNA probes(B)

Fig.2 UV-Vis spectra of AuNPs and AuNPs probesa. AuNPs; b. signal probe; c. cTnⅠ detection probe;d. HFABP detection probe; e. MYO detection probe.

Fig.3 Electrophoresis results of AuNPs before and after modificationLane 1: HFABP detection probe; lane 2: cTnⅠ detection probe; lane 3: MYO detection probe; lane 4: AuNPs; lane 5: signal probe; lane 6: DNA probe(25 nt); lane 7: DNA ladder(20 bp).

Fig.4 Optimization results for the amount of detection probe(A) and signal probe(B)

Fig.5 Detection results of protein chips before and after amplification(A1—A3) Detection results of signal AuNPs probe; (B1—B3) detection results of double AuNPs probes hybridization.(A1), (B1) blank control; (A2), (B2) 1 ng/mL; (A3), (B3) 32 ng/mL.

Fig.6 Specificity of myocardial injury markers detected by the developed protein chip(A) Blank control; (B) specific test of HFABP; (C) specific test of cTnⅠ; (D) specific test of MYO.

Fig.7 Standard curves of HFABP(a),cTnⅠ(b) and MYO(c)R2: a. 0.995; b. 0.994; c. 0.990.

Table 1 Comparision between ECLIA and protein chip for the detection of cTnⅠ and MYO in serum

Method	AMI(n=42)				UA(n=30)				Normal control(n=30)
	cTnⅠ		MYO		cTnⅠ		MYO		cTnⅠ		MYO
	+	-	+	-	+	-	+	-	+	-	+	-
Protein chip	31	11	37	5	6	24	15	15	1	29	7	23
ECLIA kit	30	12	35	7	6	24	16	14	1	29	6	24
χ²	0.060		0.389		0		0.067		0		0.098
P	0.807		0.533		1.000		0.796		1.000		0.754

Table 2 Comparision between protein chip and ELISA for the detection of HFABP in serum

Method	AMI(n=42)		UA(n=30)		Normal control(n=30)
	HFABP		HFABP		HFABP
	+	-	+	-	+	-
Protein chip	39	3	9	21	3	27
ELISA kit	36	6	11	19	4	28
χ²	0.498		0.300		0.097
P	0.480		0.584		0.756

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