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Method for diagnosing hepatitis e virus infection and kit

A technology for hepatitis E virus and hepatitis E, which is applied in the field of immunological diagnosis and medicine, and can solve the problems of ELISA detection method, such as lower sensitivity, easy false positive or limited application

Active Publication Date: 2015-07-22
NAT INST FOR FOOD & DRUG CONTROL +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the experimental operation of RT-PCR detection method is complicated, the required experimental equipment is expensive, the requirements for sample collection and storage are high, and the experimental results are unstable, and false positives or false negatives are prone to occur, which greatly limits its use. application
The experimental operation of ELISA detection method is simple, but the existence of anti-HEV antibody (HEV-Ab) in serum affects the detection of HEV-Ag, which reduces the sensitivity of ELISA detection method
In addition, since the level of HEV-Ab in serum is constantly changing at different stages of HEV infection, the influence of HEV-Ab on the ELISA assay is also constantly changing, which makes the detection results of the ELISA assay unstable

Method used

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  • Method for diagnosing hepatitis e virus infection and kit
  • Method for diagnosing hepatitis e virus infection and kit
  • Method for diagnosing hepatitis e virus infection and kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0156] Example 1. Detection of hepatitis E virus antigen (HEV-Ag) in urine

[0157] With the informed consent of the patient, urine samples and serum samples at the same time point were collected from a 35-year-old patient with chronic hepatitis E (Baoding Infectious Disease Hospital, Hebei Province). Urine samples and serum samples were then tested for HEV antigen (ORF2 antigen), nucleic acid, IgM and IgG antibodies using commercially available kits (see above) according to the manufacturer's instructions. The levels of alanine aminotransferase and aspartate transaminase in serum samples were detected using conventional kits (Japanese Wako Pure Chemical Industries, Ltd., catalog number: KH695). Before testing, serum samples and urine samples were stored in a -80°C refrigerator. The test results are shown in Table 1.

[0158] Table 1: Test results of urine and serum samples from a patient with chronic hepatitis E

[0159]

[0160]

[0161] Note: ALT, alanine aminotran...

Embodiment 2

[0171]Example 2. Relationship between HEV antigens in urine and HEV antigens in other types of samples

[0172] In order to study the time point when HEV antigens in urine appeared after the body was infected with HEV, and the relationship between HEV antigens in urine and HEV antigens in other types of samples, a primate model of HEV infection in cynomolgus monkeys was established. A total of 5 cynomolgus monkeys (Beijing Xierxin Institute of Biological Resources) were used. Among them, the cynomolgus monkey numbered 13C07004 (hereinafter referred to as 7004) was attacked with human type 4 HEV (strain WQ); the number was 13C07008 ( Cynomolgus monkeys hereinafter referred to as 7008) were attacked with human-source type 1 HEV (strain W2-1); cynomolgus monkeys numbered 13C08007 (hereinafter referred to as 8007) were attacked with W2-4; numbered 13C01063 (hereinafter referred to as 1063) The cynomolgus monkeys were challenged with the urine of 7004 cynomolgus monkeys infected wi...

Embodiment 3

[0199] Example 3. Detection of urine samples treated with pretreatment reagents

[0200] Three HEV-Ag positive urine samples were taken and diluted 1:1 with pretreatment reagents containing phosphate, calf serum and TritonX-100 to obtain pretreated urine samples. Parallel detection of HEV-Ag was performed on pretreated urine samples and unpretreated raw urine samples. The test results are shown in Table 4.

[0201] Table 4: HEV-Ag Detection Results of Pretreated and Unpretreated Urine Samples

[0202]

[0203] The detection result of table 4 shows: (1) after processing 1# sample with pretreatment reagent, the sensitivity of the inventive method improves, and detection result changes into true positive by false negative; (2) after processing 2# with pretreatment reagent and After the 3# sample, the detection result (HEV-Ag detection value) did not change significantly; however, considering the dilution effect of the pretreatment reagent on the sample, the HEV-Ag detection ...

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Abstract

The invention discloses a method for diagnosing whether a subject is infected with a hepatitis e virus (HEV) or suffered from hepatitis e or not. The method comprises the following steps: (1) determining existence of a hepatitis e virus antigen (HEV-Ag) in a urine sample from the subject; and (2) determining whether the subject is infected with the HEV or suffered from the hepatitis e or not, wherein the existence of the HEV-Ag in the urine sample of the subject shows that the subject is infected with the HEV or suffered from the hepatitis e. The invention further discloses a kit for the method. In addition, the invention further discloses a method for screening candidate medicines capable of resisting the hepatitis e virus infection or treating the hepatitis e, and a kit for the method.

Description

technical field [0001] The invention relates to the fields of medicine, virology and immunology, especially the field of immunological diagnosis. Specifically, the present invention discloses a method for diagnosing whether a subject is infected with hepatitis E virus (Hepatitis E Virus, HEV), or suffering from hepatitis E, which includes: (1) determining the the presence of hepatitis E virus antigen (HEV-Ag) in a urine sample from the subject; and (2) determining whether the subject is infected with HEV or has hepatitis E; wherein HEV-Ag is present in the subject The presence in the urine sample of the subject indicates that the subject is infected with HEV or has hepatitis E; and a kit for use in the method. In addition, the invention also discloses a method for screening candidate drugs capable of resisting hepatitis E virus infection or treating hepatitis E, and a kit used for the method. Background technique [0002] Hepatitis E (Hepatitis E) is mainly transmitted thr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569G01N33/576
CPCG01N33/56983G01N33/576G01N2333/005G01N2800/26
Inventor 王佑春赵晨燕耿彦生乔杉黄维金徐晓莉
Owner NAT INST FOR FOOD & DRUG CONTROL
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