Homogeneous chemiluminiscence detection method

Abstract

The invention discloses a homogeneous chemiluminiscence detection method. The method comprises the following steps: detecting a detected object by adopting a reaction principle of a sandwich method; the method is characterized in that the kit for detecting a detected object is composed of nano-gold photosensitive particles and luminescent particles, and the nano-gold photosensitive particles are conjugates obtained by coupling a compound of the nano-gold particles and a photosensitizer with a compound capable of generating specific binding pairing; the light-emitting particles are conjugates of light-emitting microspheres and bioactive molecules such as antibodies. According to the invention, gold nanoparticles are used as a carrier; a water-soluble photosensitizer is electrostatically adsorbed, and energy is transferred to photosensitizer molecules combined with the water-soluble photosensitizer in a non-radiative energy transfer mode, so the water-soluble photosensitizer is obtained.The molecules reach an excited state, so triplet-state oxygen is excited to a singlet state, the singlet-state oxygen content is increased, the difference between the wavelength of excitation light and the emission wavelength is increased, the detection identification degree is improved, and the detection precision, accuracy and sensitivity are enhanced.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a homogeneous chemiluminescent detection method. Background technique [0002] Chemiluminescence is currently the most advanced immunoassay methodology. According to different markers, it can be divided into direct chemiluminescence, enzymatic chemiluminescence and electrochemiluminescence; according to whether there is a separation and cleaning step, it can be divided into heterogeneous chemiluminescence and Homogeneous Chemiluminescence. At present, in terms of in vitro diagnostic testing for human medicine, there are many detection products using direct chemiluminescence method and enzymatic chemiluminescence method at home and abroad, all of which are heterogeneous chemiluminescence methods. Foreign manufacturers include Abbott, Siemens, Sorin, Beckman, and Hismicom, and domestic manufacturers include New Industry, Antu, Mike, Mindray, Zecheng, and Changguang Huayi. The instrum...

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Problems solved by technology

[0003] The existing light-induced chemiluminescence method requires the use of non-water-soluble photosensitizers. The singlet oxygen quantum yield of unmodified photosensitizers is low, which affects the detection performance of the reagents; and the chemical synthesis of derivatized photosensitizers is difficult and complicated. In addition, the photosensitizer used in the existing photoluminescence system needs to be embedded in polystyrene microspheres in the organic phase, which needs to go through complicated purification methods such as centrifugation and ultrafiltration, and needs to be restored to the aqueous phase matrix , the photosensitizer is prone to loss during embedding, purification and other processing processes, which reduces the photosensitivity effect, and the existing photochemiluminescence technology has little difference between the excitation wavelength and the detection wavelength, which is easily disturbed and greatly affected by the fluorescence background

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