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Detection method of trace hemp in saliva sample and used SPR (surface plasmon resonance)chip

A detection method and a technology in a sample are applied in the detection field to achieve the effects of rapid detection, convenient sampling and high sensitivity

Inactive Publication Date: 2017-10-10
浙江诺迦生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Currently, there is no report on the rapid detection of marijuana using surface plasmon resonance technology

Method used

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  • Detection method of trace hemp in saliva sample and used SPR (surface plasmon resonance)chip
  • Detection method of trace hemp in saliva sample and used SPR (surface plasmon resonance)chip

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Embodiment 1

[0042] THC is coupled to protein carriers KLH and OVA respectively through phenolic hydroxyl groups to obtain THC-KLH and THC-OVA.

[0043] 15 BALB / C mice were immunized with THC-KLH and the fast immune adjuvant QuickAntibody-Mouse5W, and the antiserum titers of the immunized mice against methamphetamine were detected with THC-OVA after 5 weeks; 5 mice with affinity antibody (antiserum ELISA titer above 6K).

[0044] B lymphocytes from the spleen were fused with mouse myeloma cells SP2 / 0, and high-specificity and high-affinity anti-methamphetamine antibody hybridoma cell lines were obtained through monoclonal screening. ELISA titers were 100 above 10K. Then, THC-OVA was spotted and coupled to the SPRi carboxyl chip, and the amounts of THC-OVA were 30ng, 3ng, 0.3ng, and 0.03ng respectively; in addition, OVA was also spotted as a negative control. The surface plasmon resonance SPRi high-throughput biomolecular interaction instrument was used to analyze the binding reaction betw...

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Abstract

The invention relates to a detection method of trace hemp in a saliva sample and an SPR (surface plasmon resonance) chip used in the method. According to the detection method, the THC (tetrahydrocannabinol) content of the to-be-detected sample is detected on an SPR instrument by use of the SPR chip, the SPR chip is an anti-THC antibody SPR chip, and the anti-THC antibody SPR chip is prepared by coating a 3D epoxy SPR chip with an anti-THC Fab segment. The prepared SPR chip is a specific antibody coupled with anti-THC on the surface, can specifically combine THC molecules in complex samples and thus can be applied to detection of trace hemp in the saliva sample. Meanwhile, when the prepared anti-THC antibody SPR chip is used for detecting THC in the saliva sample of a hemp taking person on the SPR instrument, the quick, accurate and sensitive effect can be realized, the detection sensitivity can at least reach 0.3-3 ng / mL, and the requirement of quick detection of crime scenes can be met.

Description

technical field [0001] The invention relates to a detection method of drug marijuana, in particular to a detection method of a trace amount of marijuana in a saliva sample and an SPR chip used therefor, belonging to the technical field of detection. [0002] The meaning of following expression among the present invention is: [0003] THC: Tetrahydrocannabinol [0004] KLH: Hemocyanin [0005] OVA: chicken ovalbumin Background technique [0006] Marijuana, whose main effective chemical component is THC, has mental and physiological active effects after smoking or oral administration. It is considered to be one of the world's three major drugs, and its social hazards are very serious. [0007] Currently, testing methods for cannabis mainly include conventional chemical methods, capillary electrophoresis, thin-layer chromatography, high-performance liquid chromatography, gas chromatography and gas chromatography / mass spectrometry, immunoassays, and biological assays. Among ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/94G01N33/543G01N21/552C07K16/44
CPCC07K16/44G01N21/553G01N33/54393G01N33/948
Inventor 范春雷程向荣陈喆田男
Owner 浙江诺迦生物科技有限公司
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