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Fast detecting and source-tracing method for cryptosporidium and giardia lamblia in urban sewage

A technology for cryptosporidium and urban sewage, which is applied to the rapid detection and traceability of giardia, and the field of cryptosporidium in urban sewage, which can solve the problems of high requirements, complicated operation and high cost, so as to improve the detection level and overcome the demand Great, easy-to-operate effect

Inactive Publication Date: 2017-09-19
EAST CHINA UNIV OF SCI & TECH
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  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to provide a rapid detection and traceability method of Cryptosporidium and Giardia in urban sewage, so as to solve the problems of large sample size, complicated operation, high requirements, large errors, and ineffective species traceability in existing detection technologies. Ideal and costly issues

Method used

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  • Fast detecting and source-tracing method for cryptosporidium and giardia lamblia in urban sewage
  • Fast detecting and source-tracing method for cryptosporidium and giardia lamblia in urban sewage
  • Fast detecting and source-tracing method for cryptosporidium and giardia lamblia in urban sewage

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Embodiment 1

[0057] Sewage samples were collected from the sewage pump station, and after the above steps of centrifugation, DNA extraction, PCR, etc., positive DNA samples of Cryptosporidium in cow dung (collected from a dairy farm in Xinghua, Jiangsu) were used as positive controls to test the cryptosporidium in sewage. sporozoites were detected.

[0058] The result is as figure 1 As shown, the amplified products of samples 1-4, 8, 11-12, 15, and 26 are about 580 bp, which is the 18S rRNA gene of Cryptosporidium, so it shows that these samples contain Cryptosporidium, while other samples do not. Contains Cryptosporidium.

Embodiment 2

[0060] Carry out Giardia detection with the DNA sample among the embodiment 1, use the Giardia positive DNA sample in the cow dung (gathered from a dairy farm in Xinghua, Jiangsu with Example 1) as positive control, to the Giardia in the sewage Insect detection.

[0061] The result is as figure 2 As shown, the amplified products of samples 1-4, 8-9, 11-15, 19-20, 27, 30, 35-36 are 530bp, which is the tpi gene of Giardia, so it shows that these samples contain Giardia worms, while the other samples did not contain Giardia.

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Abstract

The invention provides a fast detecting and source-tracing method for cryptosporidium and giardia lamblia in urban sewage. The method comprises the following steps of: 1) collecting a proper amount of urban sewage, leaving the urban sewage for a while, abandoning sewage in the upper layer and collecting sewage in the lower layer; 2) repeating the step 1) to a certain water sample total amount; 3) centrifugalizing the collected water sample and collecting a precipitate; 4) extracting DNA in the precipitate; 5) performing PCR and sequencing on the extracted DNA to detect cryptosporidium and giardia lamblia in the urban sewage; and 6) comparing the sequence obtained in the step 5) with a sequence in the GenBank to determine the pest species or genotypes. By virtue of a method of performing PCR on the sewage after centrifugal precipitation, the invention provides the fast detecting and source-tracing method for cryptosporidium and giardia lamblia in the urban sewage, wherein the method is small in demand in sample volume, simple to operate, accurate in result, ideal in species-fixing and source-tracing effect and low in cost.

Description

technical field [0001] The invention relates to the field of sewage detection, in particular to a rapid detection and traceability method for Cryptosporidium and Giardia in urban sewage. Background technique [0002] Cryptosporidium and Giardia are common apicomplexan parasitic protozoa with strong host adaptability, can infect humans and various animals, and cause symptoms such as diarrhea and vomiting in the host. For immunocompromised populations such as AIDS patients and organ transplant patients, infection with Cryptosporidium can even be life-threatening. Cryptosporidium and Giardia have had many outbreaks around the world, causing more than 400,000 infections and more than 100 deaths in the most serious cases, seriously endangering public health security. [0003] According to the "Drinking Water Sanitation Standard" (GB5749-2006) revised and completed at the end of 2006 in my country, Cryptosporidium and Giardia are the only two protozoa included in the control indi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12Q1/04
CPCC12Q1/6869C12Q2549/119C12Q2535/122Y02A50/30
Inventor 黄成琛胡越王琳王元非李娜郭亚琼冯耀宇肖立华
Owner EAST CHINA UNIV OF SCI & TECH
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