CN102184350A - Method for building population genetic resource bank - Google Patents
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Abstract
The invention belongs to the field of building of population genetic resource bank, and provides a method for scientifically building a population genetic resource bank, in particular relating to a scheme for building a population genetic resource bank for seventeen native ethnic minorities in Guizhou province. The genetic resource bank comprises an information database, a plasma specimen library, a total RNA (ribonucleic acid) spare library and a DNA (deoxyribonucleic acid) specimen library. The genetic resource bank has the following characteristics: 1, the genetic resource bank is large in specimen mount and diversified in structure, and can meet the requirements of different research strategies on specimens; 2, the genetic resource bank is complete in materials and available for continuing study: through carrying out detailed demographic, genetic and epidemiological data recording on each object and carrying out clinical examination, serologic tests for hepatitis B and various blood biochemical and immune tests, the information database of a system is established; 3, a normalized acquisition, storage and management system is established; and 4, a normal complete genetic resource and specimen collection method (comprising the steps such as propaganda and mobilization, investigation and sampling, clinical examination, DNA extraction, and the like) and data input and laboratory management methods are established.
Description
Technical field
The invention belongs to the structure field of population genetic resources bank, be specifically related to a kind of method that 17 generation in Guizhou Province occupy ethnic group's genetic resource that makes up.
Background technology
Along with finishing and the progress of full genome association study work of the Human Genome Project, genomic variation or polymorphism have become possibility between deep comprehensively understanding individuality and colony, and demonstrate importance day by day in many-sides such as research complex disease related gene, Population Genetics, anthropology, origin and evolution.Mankind's inheritance resources is not only the basic material of carrying out the human genome bio-diversity, understanding descent of man and evolution, also is the material base of research human genetic disease and many major diseases.
China is a populous nation, and the Chinese nation not only accounts for 22% of world population sum, and is a multi-nationality country, except that Han nationality, also has 55 ethnic groups that discerned, is the abundantest country of mankind's inheritance resources.Guizhou Province is a multi-national province, and 49 national compositions are arranged, and generation occupies 17 of ethnic groups, and wherein Miao ethnic group's population accounts for 49.8% of national Miao ethnic group total population; Bouyei's population accounts for 97.3% of national Bouyei total population; The Dong nationality's population accounts for 55.7% of national the Dong nationality total population; Gelao's population accounts for 98.2% of national Gelao total population; Shui nationality population accounts for 90.86% of national Shui nationality total population, the Guizhou minority nationality composition occupies the 5th in the whole nation, the population of ethnic minorities occupy the 3rd in the whole nation, constituted extremely precious genetic resources, especially the national population of some remote districts is because the restriction in geographic position, the extroversion of gene flows slower, has formed the special genetic resources isolated population and the extended familys of genetic disease.This class nationality has the single relatively characteristics of genetic background, thereby has intactly kept some rare inhereditary feature and disease gene, is resource rare in the gene studies.Fast development along with economy, traffic, information; the change of life style; these resources are being faced with the danger of running off and mixing; in order to protect these precious local nationalities' genetic resources; better carry out researchs such as Population Genetics and disease related gene, having set up Guizhou generation, to occupy ethnic group's genetic resources gene pool extremely urgent.
Though domestic existing research institution has set up all kinds of population genetic resources banks at present, all there are situations such as each population sample amount is few, sample collection shortage systematicness, science to some extent in these resources banks.The mentioned Guizhou Province's generation of the present invention occupies ethnic group's genetic resource and has then that sample size is big, the diversified characteristics of structure, each national sample size is greater than 500 parts, have national even above 1000 parts, 10018 parts of resources bank sample totals, can satisfy of the requirement of different research strategies, better provide basic resource for follow-up multiclass experimental study to sample.
The methodological science of structure population genetic resources bank provided by the present invention is effective: questionnaire design system, full and accurate; Sample collection method safety, reliable; Clinical health check-up and all kinds of biochemistry, immunologic test system and pointed; Sample preservation and management regulation, reasonable.The constructed genetic resource that goes out of method provided by the invention can satisfy the multiclass experimental study and continue research, for the research of aspects such as follow-up disease related gene, Population Genetics provides research material more full and accurate, science.
Summary of the invention
The objective of the invention is to, a kind of method that scientifically makes up the population genetic resources bank is provided, provide 17 generation in Guizhou Province to occupy the constructing plan of ethnic group's genetic resource.
The inventor has built up 17 generation in Guizhou with considerable scale, all information, standardized management and has occupied ethnic group's genetic resource, comprising information database, plasma sample storehouse, total standby storehouse of RNA and dna sample storehouse.The inventor gathers Guizhou Province generation altogether and occupies totally 10018 parts of 17 ethnic group's isolated crowds and Han nationality's isolated crowd blood samples, finished the extracting genome DNA of sample, and tentatively set up Guizhou minority nationality physique epidemic data storehouse, preserved the genetic resources that 10018 parts of Guizhou 17 generation occupy the crowd of ethnic group corresponding D NA, blood plasma and clinical data.
This genetic resource has following characteristics: 1, sample size is big, structure is diversified, can satisfy of the requirement of different research strategies: guarantee that all there is bigger sample size in each nationality of being gathered, and can satisfy the correlative study of follow-up and multiple disease in conjunction with clinical data and physical examination result substantially to sample.2, all information, can be for continuing research: to every object all detail record demography, hereditary disease and epidemiologic data, and carried out clinical health check-up, the detection of hepatitis B serology and every blood biochemical and immunologic test, set up the information database of system.3, normalized collection, preservation and management system.4, set up standard from publicity and mobilization, investigation sampling, clinical health check-up, the genetic resources that a cover such as DNA extraction is complete and the method for specimen collection and data loading, laboratory room managing way.
Technical scheme of the present invention: a kind of method that makes up the population genetic resources bank.It is as follows to make up the genetic resource concrete steps:
(1), preparation for acquiring object inhabitation location message summary table, questionnaire, health check-up registration form, sample collection consumptive material, clinical health check-up equipment and clinical biochemical detecting instrument, staff's grouping.
(2), the researchist investigates and fills in acquisition target inhabitation location message summary table, tissue is sampled the crowd and accepts survey and sign genetic resources investigation Informed Consent Form to the corresponding location simultaneously: be sampled the people under normal, the state of mind of regaining consciousness, read over Informed Consent Form, agree the back signature; Or having under witness's the situation, the content of Informed Consent Form is read to being sampled the people listened, be sampled and sign in person or put one's fingerprint after the people agrees; By registration group researchist questionnaire table and Informed Consent Form are numbered simultaneously, fill in and be sampled people's basic document and survey result, and gather hair and nail.
(3), health check-up group researchist carries out information gathering of science of heredity sign and health examination to being sampled the crowd: carry out detailed health check-up to being sampled object, and fill in the health check-up registration form, health check-up is carried out diagnosis and treatment to the people that is sampled that difficult and complicated illness is arranged after finishing.
(4), blood sampling group researchist takes a blood sample to being sampled the crowd, heparin tube is numbered and the registration related content: each is sampled object and uses EDTA.Na respectively
2Anticoagulant tube and the blood sampling of sodium citrate anticoagulant tube, each manages equal venous blood samples 3m1.
(5), meal missing allowance granting and receipts table group researchist check UP registration form inspection, replenish and reclaim the granting of meal missing allowance.
(6), experiment processed group researchist in time gets EDTA.Na
2It is an amount of that the anticoagulant tube blood sample adds the RNA protective agent in right amount, and-80 ℃ of cryopreservation extract in order to follow-up total RNA.
(7), experiment processed group researchist centrifuging EDTA.Na
2Blood plasma in the anticoagulant tube blood sample and haemocyte, and haemocyte in the separate blood partly carried out the extraction of genomic DNA and quantitatively.
(8), the EDTA.Na of blood screening group researchist to having separated
2Blood plasma partly carries out all kinds of clinical biochemicals and immune indexes detection (as the examination of hepatitis B serology, glucose-6-phosphate dehydrogenase (G6PD) examination, β-thalassemia examination, the rheumatism factor, alpha-fetoprotein detection, various enzymatic activity detections etc.) in the anticoagulant tube blood sample; In time sodium citrate anticoagulant tube blood sample is carried out blood group evaluation and routine blood test detection simultaneously.
(9), experiment processed group researchist reaches quantitatively the extraction that blood sample in the sodium citrate anticoagulant tube carries out genomic DNA.
(10), set up 17 generation in Guizhou and occupy ethnic group genetic resources dna sample storehouse: put into-80 ℃ of ultra low temperature freezers after the sample genomic dna that will test processed group researchist extracting is checked digit and preserve.
(11), set up 17 generation in Guizhou and occupy ethnic group genetic resources plasma sample storehouse: EDTA.Na
2Blood plasma in the anticoagulant tube blood sample is put into-80 ℃ of ultra low temperature freezers and is preserved after partly checking digit.
(12), set up 17 generation in Guizhou Province and occupy the standby storehouse of the total RNA of ethnic group's genetic resources: will test the processed group researchist and add the protectant EDTA.Na of RNA
2The anticoagulant tube blood sample places-80 ℃ of ultra low temperature freezers to preserve, and the total RNA when needing in order to follow-up study extracts, and carries out the mRNA expression study.
(13), setting up 17 generation in Guizhou and occupy ethnic group's genetic resources information database: files in the original written information numbering of genetic resources bookbinding back, information material deposits computer in, set up the full and accurate information database of system with database software, the preservation of imprinting CDs simultaneously, and be responsible for the record management work of database by full-time researchist.
(14), formulate " 17 generation in Guizhou occupy ethnic group's genetic resource and make up guide " and " 17 generation in Guizhou occupy ethnic group's genetic resource standardized management and sample preservation, receive rules ": set up standard from publicity and mobilization, the investigation sampling, clinical health check-up, the genetic resources that a cover such as DNA extraction is complete and the method for specimen collection and data loading, laboratory room managing way.
The staff is divided into registration group, health check-up group, blood sampling group, meal missing allowance granting and receipts table group, blood screening group, experiment processed group in the abovementioned steps (1).Acquisition target inhabitation location message summary table content comprises the investigation to acquisition target inhabitation location physical geography information, economic geography information, medical geography information and Social Culture information.
Questionnaire is mainly demography, hereditary disease, epidemiology content in the step (2), comprises the investigation of family, present illness history, passing medical history etc.; Hair of gathering and nail splitting are preserved for future use.
The middle science of heredity sign information gathering project of step (3) comprises in the step (3): span, sitting height, pelvis is wide, tragus point is high, head length, the volume minimum is wide, wide between angle of mandible, ose breadth, head height on the ear, the form surface height, bilabial, physiognomic ear breadth, Darwin's knot, the ear-lobe shape, Mongolia's pleat, skin of upper lip portion height, the colour of skin, hair, the tuorbillion direction, the tuorbillion number, the tuorbillion position, the face type, the antimongoloid slant degree, the total form of nose, the nose substrate, nasion height, the nose direction, nostril form, shoulder breadth, tranquil chest measurement, waistline, the head breadth, face width, wide between angulus oculi medialis, mouth is wide, appearance face height, nasal height, physiognomic ear length, level is enclosed, the auricle profile, the upper eyelid gauffer, the upper lip bulging, red lip thickness, beard, the palpebral fissure height, fingerprint, sour jujube point is high before the bone.Assembled item comprises: span, sitting height, pelvis is wide, tragus point is high, head length, the volume minimum is wide, wide between angle of mandible, ose breadth, head height on the ear, the form surface height, bilabial, physiognomic ear breadth, Darwin's knot, the ear-lobe shape, Mongolia's pleat, skin of upper lip portion height, the colour of skin, hair, the tuorbillion direction, the tuorbillion number, the tuorbillion position, the face type, the antimongoloid slant degree, the total form of nose, the nose substrate, nasion height, the nose direction, nostril form, shoulder breadth, tranquil chest measurement, waistline, the head breadth, face width, wide between angulus oculi medialis, mouth is wide, appearance face height, nasal height, physiognomic ear length, level is enclosed, the auricle profile, the upper eyelid gauffer, the upper lip bulging, red lip thickness, beard, the palpebral fissure height, fingerprint, sour jujube point is high before the bone.The health check-up project is in the step (3): generalized case: height, body weight, blood pressure, and cooperate on-the-spot physical examination, understand the difference problem of health; 5 of internal medicine: the heart, lung, liver, spleen, nervous system, by physical examination, understand the sign that has or not internal disease or internal disease; Surgery: skin, backbone, four limbs, thyroid gland, by palpation and physical examination, understand the basic condition of surgery system; Ophthalmology: eyesight, color discrimination, external eyes, intraocular pressure, eyeground, slit lamp examination, whether understanding eyeground, blood vessel have pathology; The department of stomatology: check under lip, cheek, tooth, gums, periodontal, tongue, palate, the parotid gland, salivary gland, the temporo,, understand the sign that has or not oral cavity department disease or oral cavity department disease by physical examination; Hals,Nasen und Ohrenheilkunde: diaphragm, pharyngeal, throat in hearing, external ear, inner ear, nasal cavity, the nose, understand the ear,nose ﹠ throat basic condition.
EDTA.Na in the step (4)
2Each vein blood volume that extracts is 3ml in anticoagulant tube and the sodium citrate anticoagulant tube.
The RNA protective agent that adds in the step (6) is TRIzol, gets EDTA.Na
2Anticoagulant tube blood 500ul adds RNA protective agent TRIzol 1000ul, and the total RNA when needing in order to follow-up study extracts, and carries out the expression study of mRNA.
The extracting method that genomic DNA is adopted in step (7) and (9) is the phenol-chloroform extraction process.
EDTA.Na to having separated in the step (8)
2Clinical biochemical that blood plasma partly carries out in the anticoagulant tube blood sample and immune indexes test item are the examination of hepatitis B serology, the rheumatism factor, alpha-fetoprotein detection, the detection of various enzymatic activity, glucose-6-phosphate dehydrogenase (G6PD) examination, β-thalassemia examination etc.
The sample store method is-80 ℃ of ultra low temperature freezers preservations in step (10), (11), (12).
To adopt software be Access software in the typing of genetic resources information in the step (13), detail record every be sampled demography in the object questionnaire table, hereditary disease, epidemiologic data data and science of heredity sign information separately, clinical health check-up, the detection of hepatitis B serology, every blood biochemical and immunologic test result and genomic DNA quantitative data.
The above make up that 17 generation in Guizhou occupy the method for ethnic group's genetic resource and when making up other population genetic resource databases to the employing of this construction method.
Description of drawings
Fig. 1 is the healthy and genetic resources investigation Informed Consent Form of isolated crowd.
Fig. 2 and Fig. 3 are the healthy and genetic resources questionnaires of isolated crowd.
Fig. 4 is the healthy and genetic resources investigation health check-up registration form of isolated crowd.
Fig. 5 and Fig. 6 are that 17 generation in Guizhou occupy ethnic group's genetic resource database diagram; Figure is the genetic resources information data sectional drawing under the Access software administration.
Embodiment
Make up Guizhou generation below in conjunction with the inventor and occupy the embodiment of 17 ethnic group's genetic resources the present invention is described in further detail, but not as limitation of the present invention.
Embodiment: the structure that 17 generation in Guizhou occupy ethnic group's genetic resource, carry out according to the following steps:
(1), preparation for acquiring object inhabitation location message summary table, questionnaire, health check-up registration form, sample collection consumptive material, clinical health check-up equipment, clinical biochemical detecting instrument, carry out staff's grouping:
The sample collection consumptive material of preparing, clinical health check-up equipment and clinical biochemical detecting instrument have: questionnaire, the health check-up registration form, disposable blood taking needle, disposable heparin tube (band anti-coagulants), disposable heparin tube (no anti-coagulants), medicinal alcohol, the tincture of iodine, cotton swab, medicine bag, ice bag, high-quality waterproof marking pen, ball pen, scissors, nail clipper, tablecloth, rag, toiler roll, refuse bag, old newspaper, letter is signed paper, soft transcript, the large size blank sheet of paper, paste, pesticide, sanitizer.
The clinical health check-up equipment of preparing has: height and weight claims, sphygmomanometer, stethoscope, colour blindness and anomalous trichromatism detection table.
The clinical biochemical detecting instrument of preparing has: optical microscope, self-poise low speed centrifuge, 721 spectrophotometers (supporting cuvette), thermostat water bath; Screening agent: erythrocyte fragility reagent, G6PD reagent, content of hemoglobin reagent; Glass test tube, reagent bottle, distilled water, Eppendorf pipe, 96 hole ELISA Plate; The trace adjustable pipette, suction is chewed, glass is adjustable application of sample bottle; Blood count type reagent: anti-A ABS, microslide and cotton swab rod.
The researchist is divided into registration group, health check-up group, blood sampling group, meal missing allowance granting and receipts table group, blood screening group, six groups of experiment processed group.
(2), researchist investigation and fill in acquisition target inhabitation location message summary table, tissue is sampled the crowd and accepts survey to the corresponding location and sign genetic resources and investigate Informed Consent Form simultaneously:
By the investigator live on-site physical geography information, economic geography information, medical geography information and Social Culture information of acquisition target is investigated, and filled in acquisition target inhabitation location message summary table content.Being sampled the crowd by local government and hygiene department's tissue simultaneously participates in healthy and the genetic resources investigation to the corresponding location, each department, explain this investigation and inform the investigation step to the masses by registration group researchist, be sampled the people under normal, the clear-headed state of mind, read over Informed Consent Form, agree the back signature; Or having under witness's the situation, the content of Informed Consent Form is read to being sampled the people listened, be sampled and sign in person or put one's fingerprint after the people agrees.
Registration group researchist fills in and is sampled people's basic document questionnaire and Informed Consent Form numbering, carries out survey and record according to the questionnaire content to being sampled the crowd.Questionnaire is mainly demography, hereditary disease, epidemiology content, comprises the investigation of family, present illness history, passing medical history etc.
Gather hair and nail after survey, splitting is preserved for future use.
(3), health check-up group researchist is to being sampled collection and the health examination that the crowd carries out science of heredity sign information:
Health check-up group researchist is to being sampled the collection that object carries out science of heredity sign information, and the measurement project comprises: span, sitting height, pelvis is wide, tragus point is high, head length, the volume minimum is wide, wide between angle of mandible, ose breadth, head height on the ear, the form surface height, bilabial, physiognomic ear breadth, Darwin's knot, the ear-lobe shape, Mongolia's pleat, skin of upper lip portion height, the colour of skin, hair, the tuorbillion direction, the tuorbillion number, the tuorbillion position, the face type, the antimongoloid slant degree, the total form of nose, the nose substrate, nasion height, the nose direction, nostril form, shoulder breadth, tranquil chest measurement, waistline, the head breadth, face width, wide between angulus oculi medialis, mouth is wide, appearance face height, nasal height, physiognomic ear length, level is enclosed, the auricle profile, the upper eyelid gauffer, the upper lip bulging, red lip thickness, beard, the palpebral fissure height, fingerprint, sour jujube point is high before the bone.
Health check-up group researchist carries out detailed health check-up to being sampled object, and fills in the health check-up registration form, and the health check-up project is: generalized case: height, body weight, blood pressure, and cooperate on-the-spot physical examination, understand the difference problem of health; 5 of internal medicine: the heart, lung, liver, spleen, nervous system, by physical examination, understand the sign that has or not internal disease or internal disease; Surgery: skin, backbone, four limbs, thyroid gland, by palpation and physical examination, understand the basic condition of surgery system; Ophthalmology: eyesight, color discrimination, external eyes, intraocular pressure, eyeground, slit lamp examination, whether understanding eyeground, blood vessel have pathology; The department of stomatology: check under lip, cheek, tooth, gums, periodontal, tongue, palate, the parotid gland, salivary gland, the temporo,, understand the sign that has or not oral cavity department disease or oral cavity department disease by physical examination; Hals,Nasen und Ohrenheilkunde: diaphragm, pharyngeal, throat in hearing, external ear, inner ear, nasal cavity, the nose, understand the ear,nose ﹠ throat basic condition.
After health check-up finished, health check-up group doctor was sampled people's diagnosis and treatment of being correlated with to what difficult and complicated illness arranged.
(4), blood sampling group researchist takes a blood sample to being sampled the crowd, heparin tube is numbered and the registration related content:
Each is sampled object and uses EDTA.Na respectively
2Anticoagulant tube and the blood sampling of sodium citrate anticoagulant tube, each manages equal venous blood samples 3ml.
(5), meal missing allowance granting and receipts table group researchist check UP registration form inspection, replenish and reclaim, provide meal missing allowance simultaneously.
(6), experiment processed group researchist in time gets EDTA.Na
2Anticoagulant tube blood sample 500ul adds RNA protective agent TRIzol 1000ul, and-80 ℃ of cryopreservation extract in order to follow-up total RNA, carry out the expression study of mRNA.
(7), EDTA.Na
2The extraction of the separation of anticoagulant tube blood constituent and blood sample genomic DNA and quantitative:
Experiment processed group researchist centrifuging EDTA.Na
2Blood plasma in the anticoagulant tube blood sample and haemocyte partly adopt the phenol-chloroform extraction process to carry out the extraction of genomic DNA to haemocyte in the separate blood sample again, adopt U.S. Beckman DU 640 protein core acid cut component analysis instrument carry out DNA quantitatively.
(8), blood sample blood group evaluation, routine blood test detection and every clinical blood biochemistry, amynologic index detect:
The EDTA.Na of blood screening group researchist to having separated
2Blood plasma partly carries out the detection of all kinds of clinical biochemicals and immune indexes in the anticoagulant tube blood sample: the examination of hepatitis B serology, glucose-6-phosphate dehydrogenase (G6PD) examination, β-thalassemia examination, the rheumatism factor, alpha-fetoprotein detection, various enzymatic activity detections etc.; In time sodium citrate anticoagulant tube blood sample is carried out blood group evaluation and routine blood test detection simultaneously.
(9), experiment processed group researchist reaches quantitatively the extraction that blood sample in the sodium citrate anticoagulant tube carries out genomic DNA.
(10), set up 17 generation in Guizhou and occupy ethnic group genetic resources dna sample storehouse: put into ad hoc-80 ℃ of dna sample storehouse ultra low temperature freezers after the sample genomic dna that will test processed group researchist extracting is checked digit and preserve (2 parts of genomic DNA samples of each research object).
(11), set up 17 generation in Guizhou and occupy ethnic group genetic resources plasma sample storehouse: EDTA.Na
2Putting into ad hoc-80 ℃ of plasma sample storehouse ultra low temperature freezers after blood plasma in the anticoagulant tube blood sample is partly checked digit preserves.
(12), set up 17 generation in Guizhou Province and occupy the standby storehouse of the total RNA of ethnic group's genetic resources: will test the processed group researchist and add the protectant EDTA.Na of RNA
2The anticoagulant tube blood sample places-80 ℃ of ultra low temperature freezers to preserve, and the total RNA when needing in order to follow-up study extracts, and carries out the mRNA expression study.
(13), setting up 17 generation in Guizhou and occupy ethnic group's genetic resources information database: files in the original written information numbering of genetic resources bookbinding back; Information material deposits computer in, adopt Access software typing genetic resources information, every of detail record is sampled demography in the object questionnaire table, hereditary disease, epidemiologic data data and science of heredity sign information separately, clinical health check-up, the detection of hepatitis B serology, every blood biochemical and immunologic test result and genomic DNA quantitative data, the information database that the system that sets up is full and accurate, the preservation of imprinting CDs simultaneously, and be responsible for the record management work of database by full-time researchist.
(14), formulate " 17 generation in Guizhou occupy ethnic group's genetic resource and make up guide " and " 17 generation in Guizhou occupy ethnic group's genetic resource standardized management and sample preservation, receive rules ": set up standard from publicity and mobilization, the investigation sampling, clinical health check-up, the genetic resources that a cover such as DNA extraction is complete and the method for specimen collection and data loading, laboratory room managing way.
Claims (12)
1. a method that makes up the population genetic resources bank relates to the structure that 17 generation in Guizhou Province occupy ethnic group's genetic resource, it is characterized in that: make up genetic resource and may further comprise the steps:
(1), preparation for acquiring object inhabitation location message summary table, questionnaire, health check-up registration form, sample collection consumptive material, clinical health check-up equipment and clinical biochemical detecting instrument, staff's grouping;
(2), researchist investigation and fill in preparation for acquiring object inhabitation location message summary table, tissue is sampled the crowd and accepts survey to the corresponding location and sign genetic resources and investigate Informed Consent Form simultaneously;
(3), health check-up group researchist carries out information gathering of science of heredity sign and health examination to being sampled the crowd;
(4), blood sampling group researchist takes a blood sample to being sampled the crowd, heparin tube is numbered and the registration related content;
(5), meal missing allowance granting and receipts table group researchist check UP registration form inspection, replenish and reclaim the granting of meal missing allowance;
(6), experiment processed group researchist in time gets an amount of EDTA.Na
2The anticoagulant tube blood sample adds-80 ℃ of preservations after an amount of RNA protective agent, in order to the extraction of follow-up total RNA;
(7), experiment processed group researchist centrifuging EDTA.Na
2Blood plasma in the anticoagulant tube blood sample and haemocyte, and haemocyte in the separate blood partly carried out the extraction of genomic DNA and quantitatively;
(8), the EDTA.Na of blood screening group researchist to having separated
2Blood plasma partly carries out the detection of all kinds of clinical biochemicals and immune indexes in the anticoagulant tube blood sample; Simultaneously sodium citrate anticoagulant tube blood sample is carried out blood group evaluation and routine blood test detection;
(9), experiment processed group researchist reaches quantitatively the extraction that blood sample in the sodium citrate anticoagulant tube carries out genomic DNA;
(10), set up 17 generation in Guizhou Province and occupy ethnic group genetic resources dna sample storehouse: put into-80 ℃ of ultra low temperature freezers after the sample genomic dna that will test processed group researchist extracting is checked digit and preserve.
(11), set up 17 generation in Guizhou Province and occupy ethnic group genetic resources plasma sample storehouse: EDTA.Na
2Blood plasma in the anticoagulant tube blood sample is put into-80 ℃ of ultra low temperature freezers and is preserved after partly checking digit.
(12), set up 17 generation in Guizhou Province and occupy the standby storehouse of the total RNA of ethnic group's genetic resources: will test the processed group researchist and add the protectant EDTA.Na of RNA
2The anticoagulant tube blood sample places-80 ℃ of ultra low temperature freezers to preserve, and the total RNA when needing in order to follow-up study extracts, and carries out the mRNA expression study;
(13), setting up 17 generation in Guizhou Province and occupy ethnic group's genetic resources information database: files in the original written information numbering of genetic resources bookbinding back, information material deposits computer in, set up the full and accurate information database of system with Access software, the preservation of imprinting CDs simultaneously, and be responsible for the record management work of database by full-time researchist.
(14), formulate " 17 generation in Guizhou occupy ethnic group's genetic resource and make up guide " and " 17 generation in Guizhou occupy ethnic group's genetic resource standardized management and sample preservation, receive rules ": set up standard from publicity and mobilization, the investigation sampling, clinical health check-up, the genetic resources that a cover such as DNA extraction is complete and the method for specimen collection and data loading, laboratory room managing way.
2. occupy the method for ethnic group's genetic resource according to 17 generation in the described structure of claim 1 Guizhou Province, it is characterized in that: the staff is divided into registration group, health check-up group, blood sampling group, meal missing allowance granting and receipts table group, blood screening group, experiment processed group in the step (1).Acquisition target inhabitation location message summary table content comprises the investigation to acquisition target inhabitation location physical geography information, economic geography information, medical geography information and Social Culture information.
3. occupy the method for ethnic group's genetic resource according to 17 generation in the described structure of claim 1 Guizhou Province, it is characterized in that: the questionnaire content is demography, hereditary disease, epidemiology content in the step (2), comprises the investigation of family, present illness history, passing medical history etc.; Hair of gathering and nail splitting are preserved for future use.
4. occupy the method for ethnic group's genetic resource according to 17 generation in the described structure of claim 1 Guizhou Province, it is characterized in that: science of heredity sign information gathering project comprises in the step (3): span, sitting height, pelvis is wide, tragus point is high, head length, the volume minimum is wide, wide between angle of mandible, ose breadth, head height on the ear, the form surface height, bilabial, physiognomic ear breadth, Darwin's knot, the ear-lobe shape, Mongolia's pleat, skin of upper lip portion height, the colour of skin, hair, the tuorbillion direction, the tuorbillion number, the tuorbillion position, the face type, the antimongoloid slant degree, the total form of nose, the nose substrate, nasion height, the nose direction, nostril form, shoulder breadth, tranquil chest measurement, waistline, the head breadth, face width, wide between angulus oculi medialis, mouth is wide, appearance face height, nasal height, physiognomic ear length, level is enclosed, the auricle profile, the upper eyelid gauffer, the upper lip bulging, red lip thickness, beard, the palpebral fissure height, fingerprint, sour jujube point is high before the bone; The health check-up project is in the step (3): generalized case: height, body weight, blood pressure, and cooperate on-the-spot physical examination, understand the difference problem of health; 5 of internal medicine: the heart, lung, liver, spleen, nervous system, by physical examination, understand the sign that has or not internal disease or internal disease; Surgery: skin, backbone, four limbs, thyroid gland, by palpation and physical examination, understand the basic condition of surgery system; Ophthalmology: eyesight, color discrimination, external eyes, intraocular pressure, eyeground, slit lamp examination, whether understanding eyeground, blood vessel have pathology; The department of stomatology: check under lip, cheek, tooth, gums, periodontal, tongue, palate, the parotid gland, salivary gland, the temporo,, understand the sign that has or not oral cavity department disease or oral cavity department disease by physical examination; Hals,Nasen und Ohrenheilkunde: diaphragm, pharyngeal, throat in hearing, external ear, inner ear, nasal cavity, the nose, understand the ear,nose ﹠ throat basic condition.
5. occupy the method for ethnic group's genetic resource according to 17 generation in the described structure of claim 1 Guizhou Province, it is characterized in that: EDTA.Na in the step (4)
2Each vein blood volume that extracts is 3ml in anticoagulant tube and the sodium citrate anticoagulant tube.
6. occupy the method for ethnic group's genetic resource according to 17 generation in the described structure of claim 1 Guizhou Province, it is characterized in that: the RNA protective agent that adds in the step (6) is TRIzol, gets EDTA.Na
2Anticoagulant tube blood 500ul adds RNA protective agent TRIzol 1000ul, and the total RNA when needing in order to follow-up study extracts, and carries out the expression study of mRNA;
7. occupy the method for ethnic group's genetic resource according to 17 generation in the described structure of claim 1 Guizhou Province, it is characterized in that: the extracting method that genomic DNA is adopted in step (7) and (9) is the phenol-chloroform extraction process.
8. occupy the method for ethnic group's genetic resource according to 17 generation in the described structure of claim 1 Guizhou Province, it is characterized in that: EDTA.Na to having separated in the step (8)
2Clinical biochemical that blood plasma partly carries out in the anticoagulant tube blood sample and immune indexes test item have: the examination of hepatitis B serology, the rheumatism factor, alpha-fetoprotein detection, the detection of various enzymatic activity, glucose-6-phosphate dehydrogenase (G6PD) examination, β-thalassemia examination etc.;
9. occupy the method for ethnic group's genetic resource according to 17 generation in the described structure of claim 1 Guizhou Province, it is characterized in that: the sample store method is-80 ℃ of ultra low temperature freezers preservations in step (10), (11), (12).
10. occupy the method for ethnic group's genetic resource according to 17 generation in the described structure of claim 1 Guizhou Province, it is characterized in that: to adopt software be Access software in the typing of genetic resources information in the step (13), detail record every be sampled demography in the object questionnaire table, hereditary disease, epidemiologic data data and science of heredity sign information separately, clinical health check-up, the detection of hepatitis B serology, every blood biochemical and immunologic test result and genomic DNA quantitative data.
11. one kind obtains the method that 17 generation in Guizhou Province occupy ethnic group's genetic resource by making up resource, it is characterized in that: successfully made up and comprise the Pu loud, high-pitched sound Yi nationality, distributed over Yunnan, Sichuan and Guizhou, the Da Zhai flower seedling, the show water Hui ethnic group, husky well pot circle seedling, the Sha Jing Gelao, greening Bai nationality, the Jin Po Manchu, the Zhong Mountain Bouyei, the Zhong Mountain Yi nationality, distributed over Yunnan, Sichuan and Guizhou, the Maonan, korneforos, beat the plain Bouyei, the luxuriant blue Bouyei, the white trousers precious jade of Yao Shan, the blue or green trousers precious jade in the precious jade foot of a hill or mountain, precious jade sorrow long gown precious jade, the three hole Shui nationality, Longjiang the Dong nationality, the flat Zhuang of silver, the husky Miao ethnic group of stone mountain, height increases the Dong nationality, Leishan longuette seedling, the Ji Dong Mulao, the six fort Shes, the west of a river Mongols, cloud house Tujia, Miao ethnic group of shuttle man, the seven tree Qiang occupy ethnic group's genetic resource and Changshun in 17 interior generation, Libo and the crowd of Bijie Prefecture Han nationality genetic resource.
12. as method or the application of step in making up other population genetic resources banks as described in each among the claim 1-11.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109493935A (en) * | 2018-12-21 | 2019-03-19 | 宜宝科技（北京）有限公司 | Inspection report management method and device based on hospital end |
| CN111008824A (en) * | 2019-12-13 | 2020-04-14 | 陇东学院 | Scientific research help-seeking management system based on multi-disciplinary fusion of network and geographic information technology |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109493935A (en) * | 2018-12-21 | 2019-03-19 | 宜宝科技（北京）有限公司 | Inspection report management method and device based on hospital end |
| CN111008824A (en) * | 2019-12-13 | 2020-04-14 | 陇东学院 | Scientific research help-seeking management system based on multi-disciplinary fusion of network and geographic information technology |
| CN111008824B (en) * | 2019-12-13 | 2023-08-18 | 陇东学院 | Scientific research help-seeking management system based on network and geographic information technology multidisciplinary fusion |
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