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HCMVPP65 antigenemia indirect immunofluorescence method detection reagent kit

A technology for detecting kits and antigenemia, applied in fluorescence/phosphorescence, measuring devices, instruments, etc., can solve problems such as complex process, low specificity and sensitivity, and unsatisfactory results

Inactive Publication Date: 2008-09-10
天津市秀鹏生物技术开发有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The original HCMVP65 antigenemia indirect immunofluorescence detection test, the production method is backward, the process is complicated, the intermediate process is difficult to control, and the result is not ideal
low specificity and sensitivity

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0079] Blood Sample Collection

[0080] Draw 3ml of venous blood and use heparin anticoagulant or EDTA anticoagulant. Gently invert to mix the sample.

[0081] Experimental procedure

[0082] ● Add 2ml whole blood + 6ml prepared erythrocyte lysate into a 15ml conical centrifuge tube. Mix gently with a pipette. Let stand at room temperature for 5 minutes. Centrifuge at +2~+8°C, 160g for 10 minutes. Gently invert the tube and pour the supernatant into a disinfectant solution (eg, 0.5% or 1% sodium hypochlorite). Keep the pellet and handle carefully, do not remove the cell pellet. Add 3ml of prepared erythrocyte lysate to resuspend the blood cells. Do not leave at room temperature, proceed to the next step immediately. Centrifuge at +2~+8°C, 160g for 10 minutes. Invert the tube again to remove the supernatant according to the previous method, and be careful not to remove the cell pellet. Add 1ml of the prepared reagent A (PBS) to resuspend the blood cells, and mix them ...

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PUM

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Abstract

The invention discloses a HCMVPP65 antigenemia indirect immune fluorescence method detection kit, which uses cytomegalovirus-AD169 virus strain pp65 protein as the immunogen to immune a Balb / c mouse. The spleen cells of the immunized mouse and the myeloma cells of the mouse which belongs to the same type with the immune mouse are conventionally integrated, by indirect ELISA screening and finite dilution cloning, the hybridoma cell lines of the mouse cytomegalovirus pp65 protein cloning antibody are obtained, and the characteristics of the hybridoma cell lines are identified by ELISA, immune fluorescence experiment and other methods; two monoclonal antibodies that stably secrete the pp65 protein are established successfully and named respectively as 1A6 and 4A8. A monoclonal antibody which differs from the former report and aims at the pp65 protein of the cytomegalovirus (CMV) is prepared and a method used for preparing erythrocyte fast pyrolysis is established; compared with other detection kits which belongs to the same kind, the detection kit of the invention is faster, simpler and more convenient and has higher specificity and sensitivity.

Description

technical field [0001] The invention belongs to and particularly relates to a detection kit for HCMVPP65 antigenemia indirect immunofluorescence method. Background technique [0002] Current status of research and development of technologies related to detection of giant cell infection in foreign countries: Although the centrifugation technology of viral capsid shedding has been widely used, it takes a long time and the results are greatly affected by the storage time of the specimen, so its clinical application value is still unclear . Serological tests can only detect positive results after 2-4 weeks of infection, and due to the influence of immunosuppressants after organ transplantation, they cannot distinguish latent infection from active infection, so its clinical application value is limited to a certain extent. Recently, it has been reported abroad that the standardized HCMV analysis method can help predict HCMV disease. Antigenemia detection technology is a faster,...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N21/64G01N33/531
Inventor 赵卫军
Owner 天津市秀鹏生物技术开发有限公司
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