A fluorescence LAMP detection method for Cryptosporidium parvum

A technology of Cryptosporidium parvum and detection method, which is applied in the direction of microorganism-based method, measurement/inspection of microorganism, biochemical equipment and method, etc., which can solve the problem of low detection rate, high detection cost and difficult and other issues to achieve the effect of reducing the cost of detection

Inactive Publication Date: 2018-11-27
NORTHWEST A & F UNIV
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Problems solved by technology

Traditional morphological microscopic observation is time-consuming and labor-intensive, and the detection rate is low, and it also requires certain professional knowledge
Although immunological methods and molecular biology methods based on PCR are sensitive and specific, and can effectively distinguish Cryptosporidium species, the detection cost is very high and requires more expensive instruments (microplate reader, fluorescence microscope, PCR instrument, gel Imaging system, fluorescent quantitative PCR instrument, etc.), therefore, it is difficult to be accepted in grassroots application promotion

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  • A fluorescence LAMP detection method for Cryptosporidium parvum
  • A fluorescence LAMP detection method for Cryptosporidium parvum
  • A fluorescence LAMP detection method for Cryptosporidium parvum

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Embodiment Construction

[0027] In order to make the object, technical solution and advantages of the present invention clearer, the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0028] The cryptosporidium parvum fluorescence LAMP detection method comprises the following steps:

[0029] 1. First, according to the reference sequence of Cryptosporidium parvum DNAJ-like protein gene (AF177278) in GenBank, the reference sequence is as follows:

[0030]

[0031] 2. Specificity of different gene loci (SSU RNA, GP60, HSP70, actin) of Cryptosporidium parvum

[0032] Sex primer screening confirmed. Primer sequences are shown in Table 1:

[0033] Table 1 Specific primers for nested PCR detection of Cryptosporidium parvum

[0034]

[0035]

[0036] 3. According to the Cry...

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Abstract

The invention provides a fluorescence LAMP detection method for Cryptosporidium parvum and the method is specific, sensitive, low in the detection cost, short in time consuming, high in efficiency, and simple in operation. An adopted technical scheme includes the following steps: (1): screening and determination of specific primers of genetic loci which are SSU RNA, COWP, actin and HSP70 of the Cryptosporidium parvum; (2): design and synthesis of fluorescence LAMP primers of Cryptosporidium parvum, optimization of a reaction condition and drafting of a standard curve and a melting curve; and (3): testing of specificity, sensitivity and repeatability and detection application for clinical samples.

Description

1. Technical field: [0001] The invention relates to a biological detection method, in particular to a simple, sensitive, specific and rapid detection method for Cryptosporidium parvum, which can be widely used in the rapid detection of Cryptosporidium parvum in humans, animals and various environments. 2. Background technology: [0002] Cryptosporidium (Cryptosporidium), as a protozoan that widely parasitizes the digestive tract of humans and various animals, mainly causes diarrhea-based syndrome in the host, resulting in malnutrition and growth retardation. As one of the zoonotic species, Cryptosporidium parvum (C.parvum) can not only infect people through animal products such as milk, meat and fur of animals, but also pollute water sources through its feces, causing serious damage to human health and Public health poses a serious threat. Studies have shown that Cryptosporidium infection can occur in hundreds of species of animals around the world, as well as in humans. W...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6844C12Q1/04C12R1/90
CPCC12Q1/6844C12Q2531/119C12Q2563/107
Inventor 赵光辉宋军科王莎莎
Owner NORTHWEST A & F UNIV
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