CN-122012745-A - Primer group, kit, method and application for detecting schistosome and oncomelania MNP (molecular weight distribution) marker loci in water environment

Abstract

The present disclosure provides a primer set, a kit, a method and an application for detecting schistosome and oncomelania MNP marker loci in water environment. The primer set comprises at least one of a1 st primer pair to a 14 th primer pair, each primer pair comprises a forward primer and a reverse primer, and the forward primer of the 1 st primer pair, the reverse primer of the 1 st primer pair to the forward primer of the 14 th primer pair and the reverse primer of the 14 th primer pair are sequentially shown as SEQ ID NO. 1 to SEQ ID NO. 28 in a sequence table. The primer group has amplification compatibility, can be amplified in a reaction system by adopting a super-multiplex PCR technology, realizes the amplification of 14 MNP (MNP) marker loci in the reaction system, fuses a second generation sequencing platform to carry out the sequence analysis of amplified products, realizes the identification of schistosoma japonicum and Hubei oncomelania by one reaction, and has comprehensive, efficient, sensitive and accurate detection effect.

Inventors

• JIANG YONGZHONG
• XIAO ZILAN
• ZHU ZHAOLU
• YUAN YI
• HU BING
• XU JINGDONG
• Yin Qiangling
• LI BO
• TU ZHEN
• WANG GANG
• TANG MIN

Assignees

• 湖北省疾病预防控制中心（湖北省预防医学科学院）
• 湖南明了科技有限公司
• 武汉明了生物科技有限公司

Dates

Publication Date

20260512

Application Date

20260330

Claims (8)

1. 1. A primer group for detecting schistosome and oncomelania MNP marker loci in water environment is characterized by comprising at least one of a 1 st primer pair to a 14 th primer pair, wherein each primer pair comprises a forward primer and a reverse primer, and the forward primer of the 1 st primer pair, the reverse primer of the 1 st primer pair and the forward primer of the 14 th primer pair and the reverse primer of the 14 th primer pair are sequentially shown as SEQ ID NO. 1 to SEQ ID NO. 28 in a sequence table.
2. 2. A kit for detecting schistosome and oncomelania MNP marker loci in an aqueous environment, comprising the primer set of claim 1.
3. 3. A method for detecting schistosome and oncomelania MNP marker loci in an aqueous environment, the method comprising: amplifying a sample to be detected by using the primer set according to claim 1 to obtain an amplified product; sequencing the amplification product by second generation high throughput to obtain sequencing data; Comparing the sequencing data with reference genomes of schistosome and oncomelania respectively to obtain a sequencing result of the sample to be tested; and when the sequencing result is the same as the sequence of the reference genome of the schistosome and the oncomelania, judging that the schistosome and the oncomelania exist in the sample to be tested.
4. 4. The application of the MNP marker locus combination for detecting the schistosome and the oncomelania in the water environment is characterized by comprising the step of using the MNP marker locus combination for detecting the schistosome and the oncomelania in the water environment, wherein the MNP marker locus combination comprises MNP-1-MNP-14, and the positions of MNP-1-MNP-14 on a reference sequence are shown in the following table: 。
5. 5. The use according to claim 4, wherein the use comprises combining the MNP marker loci for both schistosomiasis epidemic source water pathogen epidemic species tracking and geographical distribution investigation.
6. 6. The use according to claim 4, characterized in that the use comprises combining the MNP-marking sites for assessing the risk of spread of schistosomiasis in different waters.
7. 7. The use according to claim 4, wherein the schistosome is schistosoma japonicum.
8. 8. The use according to claim 4, wherein the oncomelania is Hubei oncomelania.

Description

Primer group, kit, method and application for detecting schistosome and oncomelania MNP (molecular weight distribution) marker loci in water environment Technical Field The present disclosure relates to the technical field of molecular biological diagnosis and environmental pathogen monitoring, and in particular relates to a primer set, a kit, a method and an application for detecting schistosome and oncomelania MNP marker loci in water environment. Background Schistosomiasis is an important parasitic disease of human and animals, hundreds of millions of people are threatened by the schistosomiasis worldwide, and is one of six tropical diseases determined by WHO, and is also one of important prevention and control parasitic diseases in China. The oncomelania is required to be used as an intermediate host for transmission, when the worm eggs enter water, the worm eggs develop into cercaria, the cercaria infects the oncomelania, the oncomelania continues to develop into cercaria to release water in the oncomelania, the water body at the moment is epidemic water, the oncomelania has infectivity, the existence of schistosome cercaria in the water body of an epidemic area and the distribution and the infection rate of the oncomelania in the environment are effectively monitored, and the key of evaluating the transmission risk, early warning epidemic situation and implementing accurate prevention and control is realized. Traditional schistosomiasis monitoring relies on disease detection and artificial snail detection technologies, has the problems of low sensitivity, poor efficiency, environmental pollution and the like, and the distribution area base number of unique intermediate host oncomelania of schistosomiasis is still larger, so that the monitoring technology is time-consuming and labor-consuming. The molecular biology method, especially based on polymerase chain reaction (Polymerase chain reaction, PCR) technology, improves the sensitivity and specificity of detection, but conventional PCR is designed for a single target, if schistosome, oncomelania and the infection state thereof are to be detected simultaneously, multiple independent PCR reactions are needed, so that the detection cost is high, the time is long, the operation is complex, and the omission is easy to cause due to low sample size consumption and low target content in environmental samples, therefore, the requirement of comprehensive ecological monitoring and accurate risk assessment cannot be met by single target detection. Therefore, there is a need to develop a method that can detect schistosome and oncomelania simultaneously in an aqueous environment sample at one time, with high throughput and high sensitivity. BRIEF SUMMARY OF THE PRESENT DISCLOSURE In order to solve the problems of the prior art, the embodiment of the disclosure provides a primer group, a kit, a method and application for detecting schistosome and oncomelania MNP marking sites in water environment. The technical scheme is as follows: In one aspect, the disclosure provides a primer set for detecting schistosome and oncomelania MNP marker loci in water environment, wherein the primer set comprises at least one of a1 st primer pair to a 14 th primer pair, each primer pair comprises a forward primer and a reverse primer, and the forward primer of the 1 st primer pair, the reverse primer of the 1 st primer pair to the forward primer of the 14 th primer pair and the reverse primer of the 14 th primer pair are sequentially shown as SEQ ID NO. 1 to SEQ ID NO. 28 in a sequence table. In another aspect, the present disclosure provides a kit for detecting schistosome and oncomelania MNP marker loci in an aqueous environment, the kit comprising the above primer set. In yet another aspect, the present disclosure provides a method of detecting schistosome and oncomelania MNP marker loci in an aqueous environment, the method comprising: amplifying a sample to be detected by adopting the primer group to obtain an amplified product; sequencing the amplification product by second generation high throughput to obtain sequencing data; Comparing the sequencing data with reference genomes of schistosome and oncomelania respectively to obtain a sequencing result of the sample to be tested; and when the sequencing result is the same as the sequence of the reference genome of the schistosome and the oncomelania, judging that the schistosome and the oncomelania exist in the sample to be tested. In one aspect, the disclosure provides an application of MNP marker locus combination for detecting schistosome and oncomelania in water environment, which is characterized in that the application comprises using the MNP marker locus combination for detecting schistosome and oncomelania in water environment, wherein the MNP marker locus combination comprises MNP-1-MNP-14, and the positions of MNP-1-MNP-14 on a reference sequence are shown in the following table: MNP tag number Reference sequenceMNP ori