77 results about "Alpha-thalassemia" patented technology

This invention relates to test kit and test paper with oligonucleotide probes for diagnosing alpha-thalassemia. The test paper comprises a base, and specific oligonucleotide probes immobilized on the base. The oligonucleotide probes comprise: specific oligonucleotide probes for detecting mutation of alpha-globin gene, specific oligonucleotide probes for detecting deletion of alpha-globin gene, and base sequence complementary with the above sequences. The test kit comprises specific primer for amplifying alpha-globin gene or transcript, and specific oligonucleotide probes for detecting mutation / deletion of alpha-globin gene. The test kit and the test paper have such advantages as low cost, rapid detection, and stable results. The test kit and the test paper can be used in detection of mutation / deletion of alpha-globin gene, and prenatal diagnosis of severe thalassemia (alphaTalpha) caused by point mutation, and do not have error diagnosis.

The invention discloses a thalassemia gene detection method based on a fluorescence labeling quantitation PCR (Polymerase Chain Reaction) technology, and the detection method mainly comprises the following steps of: firstly designing a general fluorescence labeling primer; then designing an amplification primer aiming at a detection site; when the amplification primer is designed, tailing one primer of an amplification primer pair, wherein the tailing sequence is consistent to the sequence of the general fluorescence labeling primer; when PCR reaction is carried out, mixing a tailed primer group and the general fluorescence labeling primer to realize the fluorescence labeling of an amplification product; and after the PCR reaction is finished, placing PCR products into a sequencer, and confirming the lengths of different PCR products through a series of internal references of known molecular weight of a result by utilizing the Genemapper function of the sequencer to judge the genetype of a detected sample. The detection method disclosed by the invention can realize the detection of different genotypes of alpha thalassemia through a single tube.

The invention relates to a kit for integrated detection of alpha and beta mutant type thalassemias, in particular to a kit for detecting alpha and beta thalassemias mutant types by using multiple asymmetric amplification and reverse dot blot hybridization techniques. The kit of the invention consists of a polymerase chain reaction (PCR) reagent, a low-density chip and a hybridizing reagent, and contains a set of specific nucleotide polymorphism probes and PCR primers for amplifying target gene in an amplification sample. The kit can be used for detecting six alpha thalassemia locus mutations and 27 beta thalassemia mutations, which are common in China, at the same time. The detection process of the kit is faster and the detection result of the kit is more accurate, so the kit is expected to be widely used in diagnosis of thalassemia in clinic and instruction on sound child rearing.

The invention provides a test kit for alpha-thalassemia gene mutations. The test kit comprises a set of primer combinations for amplifying target genes in samples. The test kit is capable of performing embryo pretransplantation detection for all alpha-thalassemia gene mutations, and is quick in testing process and accurate in testing result.

The invention provides primers, a method and a chip for detecting alpha and / or beta-thalassemia point mutation and deletion mutation based on whole-gene capture sequencing and application of such primers, such method and such chip. The primers, the method, the chip and application thereof have the advantages that through designing of capture probes, relevant genes involved in alpha-thalassemia and beta-thalassemia are enriched and all mutation information including SNP and indel in full-length sequences of genes is detected; through addition of autosome, X-chromosome and Y-chromosome regions as well as upstream and downstream regions of coded genes as references, structure variations such as SNV and CNV are detected; compared with existing various hotspot mutation site detection technologies, the method is capable of detecting hotspot mutation information as well as some rare mutations and undiscovered new mutation types to detect and analyze full-length sequence specificity of target genes, fully covers the mutation types and makes up the defect that a conventional detection method easily causes missing detection of low-frequency mutations and rare mutations greatly.

The invention belongs to the field of biochemistry, and provides a fluorescent quantitative polymerase chain reaction (PCR) kit for detecting alpha-globin gene deletion. The kit consists of a pair of specific amplified xi globin primers, a pair of specific amplified alpha1 globin primers, a pair of specific amplified alpha2 globin primers, a pair of specific amplified beta-Actin gene primers, a fluorescent probe for specifically detecting xi globin, a fluorescent probe for specifically detecting alpha1 globin, a fluorescent probe for specifically detecting alpha2 globin, a fluorescent probe for specifically detecting beta-Actin genes, a DNA polymerase and the like. The kit has good sensitivity and accuracy for detecting depletion alpha-thalassemia, has good repeatability and stability, and is totally suitable for clinical detection of alpha-thalassemia.

The invention belongs to the technical field of biology and particularly relates to a primer set and a kit for detecting rare deletion type thalassemia. The primer set and the kit can be directly used for rapidly and stably detecting domestic 11 types of known rare deletion type thalassemia. The primer set for detecting the rare deletion type thalassemia comprises a primer set A and a primer set B, wherein the primer set A comprises 12 primers for detecting alpha-thalassemia deletion gene types; the primer set B comprises 10 primers for detecting beta-thalassemia deletion gene types. The kit can adopt a multiplex Gap-PCR (Polymerase Chain Reaction) technology to detect 11 rare deletion gene types and can be used for directly detecting 11 rare deletion thalassemia gene types for one time. The kit is simple to operate and saves time and cost when being compared with a manner of combining a nested PCR with a genetic analysis in antenatal diagnosis; conditions are created for comprehensively carrying out thalassemia screening and scientific evidences are provided for thalassemia diagnosis of premarital detection, antenatal detection and fetuses of the pregnancy.

The invention relates to the field of biotechnology, and discloses a method and kit for detecting southeast Asia deletion alpha-thalassemia. The detection method comprises the following steps of: designing a pair of primers Fd5 and Rd3 and a probe Prob-D at two sides of the broken end of a southeast Asia deletion alpha-thalassemia deleted fragment, designing a reverse primer Rq5 and a probe Prob-Q at the 5' end of the deleted fragment, extracting the DNA of a sample to be detected, performing real-time fluorescence PCR (polymerase chain reaction) by taking the DNA of the sample to be detectedas a template, and then analyzing the sample to be detected according to the real-time fluorescence PCR amplification curve. The kit disclosed by the invention comprises the primers Fd5, Rd3 and Rq5 and the probes Prob-D and Prob-Q marked with different wavelength reporting fluorophores. According to the detection method disclosed by the invention, amplification and detection are simultaneously performed according to the fluorescence signal intensity, thus saving time, improving the sensitivity, and being a simple and quick detection method with high sensitivity and easiness in popularization.

The invention discloses a joint detection kit for detecting alpha,beta-thalassemia associated with mutant genes, which comprises (1) a gene chip and (2) a primer, wherein the gene chip is provided with probes, and the probes are a sequence SEQ ID Nos:1-31 and a sequence which is complementary to the sequence SEQ ID Nos:1-31; and the primer is a sequence SEQ ID Nos:32-42. The thalassemia gene detection kit provides a platform for detecting 16 mutant genes associated with alpha-thalassemia (three deletion types and two mutant types) and beta-thalassemia, can perform synchronous joint detection,improve the specificity of detection, reduce cost and shorten detection time, and has a great significance for the screening of patients suffering from thalassemia, genetic counseling and prenatal diagnosis.

The invention discloses a thalassemia gene detection method based on a high-throughput sequencing technology. The thalassemia gene detection method mainly comprises the following steps that alpha & beta-thalassemia related gene fragments are specifically amplified based on PCR amplification primers of a span breakpoint and a mutation site designed by adopting a Gap-PCR method, library establishment is conducted on PCR products, library products are subjected to high-throughput sequencing, sequencing data uses a human genome hg19 as a reference sequence, sequencing depth values of gene loci in a target area chr16: 215000-236000 are analyzed according to a sequence alignment score Q10, and alpha-thalassemia deletion types are determined according to the distribution of the sequencing depth values of different loci; meanwhile sequence alignment is performed through the target area chr16: 215000-236000 and a target area chr16: 5246400-5248600, and alpha & beta-thalassemia mutation types are determined according to the basic types of specific sites. By the adoption of the thalassemia gene detection method, simultaneous detection of 6 mutation genetypes of alpha-thalassemia and 26 mutation genetypes of beta-thalassemia can be achieved.

The invention discloses a nested asymmetric PCR reagent kit for detecting alpha 2 globin gene point mutation. The reagent kit comprises a nested asymmetric PCR amplification primer, a fluorescent probe, a quenching probe, an LADNA polymerase and the like. The alpha-thalassemia point mutation genotype of a detected sample is detected by virtue of specific amplification of five point mutation destination fragments WS, QS, CS, CD30 and CD31 of the human alpha 2 globin gene and molecular hybridization and unwinding analysis of a specific fluorescent probe. The reagent kit has good sensitivity and accuracy to wild type and mutation type of alpha-thalassemia five point mutation positions WS, QS, CS, CD30 and CD31, has good repeatability and stability, and is totally suitable for clinical detection of alpha-thalassemia point mutation.

The invention provides a kit capable of detecting four deletion alpha-thalassemia genes (-alpha<3.7>, -alpha<4.2>, --<SEA> and --<THAI>) and three non-deletion alpha-thalassemia genes (alpha<CS>alpha, alpha<QS>alpha and alpha<WS>alpha) at the same time.The kit comprises a DNA chip, a PCR solution I and a PCR solution II.The DNA chip comprises a substrate and probes fixed to the substrate.The kit has the advantages that based on the PCR-reversal point hybridization detecting principle, corresponding amplification primers and probes are designed according to the mutation or deletion loci of genotypes, biotin is used for labeling the primers, amidogen is used for labeling the probes, the DNA chip (nylon membrane) is adopted as a substrate, the probes are fixed to the nylon membrane, and PCR products amplified by the specific primers and the probes fixed to the DNA chip are subjected to hybridization and signal developing box interpretation to diagnose thalassemia.

The invention relates to a kit for detecting Southeast Asia type alpha-thalassemia and discloses a sample treating fluid of the kit. The kit is characterized in that in the sample treating fluid, the content W / V of ammonium chloride is 0.83%, the required content W / V of casein ranges from 0.2% to 0.8% and the content V / V of glycerin ranges from 4% to 6%. The kit can conveniently and effectively detect Zeta globin chains and assist in diagnosing the Southeast Asia type alpha-thalassemia.

The invention discloses a kit for detecting common deletion form alpha-thalassemia and a using method of the kit. The kit comprises a pair of primers capable of performing simultaneous amplification of a signature sequence A1 in an alpha 1 section and a signature sequence A2 in an alpha 2 section in an alpha-globin gene cluster, a pair of primers capable of performing amplification of a --<SEA> genetype in the alpha-globin gene cluster and a pair of primers capable of performing amplification of a --<THAI> genetype in the alpha-globin gene cluster as well as a fluorescence probe for specific detection of the signature sequence A1 in the alpha 1 section, a fluorescence probe for specific detection of the signature sequence A2 in the alpha 2 section, a fluorescence probe for specific detection of a --<SEA> genetype amplicon and a fluorescence probe for specific detection of a --<THAI> genetype amplicon. As for the alpha-thalassemia globin gene deletion detection, the kit provided by the invention has high sensitivity, stability, accuracy and specificity.

The invention discloses a kit for detecting common deletion form alpha-thalassemia and a using method of the kit. The kit comprises a pair of primers capable of performing simultaneous amplification of a signature sequence A1 in an alpha 1 section and a signature sequence A2 in an alpha 2 section in an alpha-globin gene cluster, a pair of primers capable of performing amplification of a --<SEA> genetype in the alpha-globin gene cluster and a pair of primers capable of performing amplification of a --<THAI> genetype in the alpha-globin gene cluster as well as a fluorescence probe for specific detection of the signature sequence A1 in the alpha 1 section, a fluorescence probe for specific detection of the signature sequence A2 in the alpha 2 section, a fluorescence probe for specific detection of a --<SEA> genetype amplicon and a fluorescence probe for specific detection of a --<THAI> genetype amplicon. As for the alpha-thalassemia globin gene deletion detection, the kit provided by the invention has high sensitivity, stability, accuracy and specificity.

The invention relates to a method for determining a fetus alpha thalassemia gene haplotype. The method comprises the following steps: establishing a third-generation sequencing library according to ablood sample, wherein the blood sample is collected from blood samples of the father and mother of the fetus; performing the third-generation sequencing for the third-generation sequencing library, and obtaining a third-generation sequencing result; determining a father and mother alpha thalassemia gene haplotype according to the third-generation sequencing result; establishing a second-generationsequencing library according to a detection sample, wherein the detection sample is collected from a peripheral blood sample of a pregnant woman; performing the second-generation sequencing for the second-generation sequencing library, and obtaining a second-generation sequencing result; and determining the fetus alpha thalassemia gene haplotype according to the second-generation sequencing result as well as the father and mother alpha thalassemia gene haplotype.

The invention relates to a nucleic acid membrane strip for non-deletion type alpha-thalassemia gene detection. The nucleic acid membrane strip comprises a substrate and specific oligonucleotide probes fixed to the substrate, wherein the sequences of the specific oligonucleotide probes are shown in SEQ ID NO: 1-6. The invention further relates to a reagent kit for non-deletion type alpha-thalassemia gene detection and a detection method. The nucleic acid membrane strip for non-deletion type alpha-thalassemia gene detection has the advantages of being high in adaptability and specificity. The reagent kit for non-deletion type alpha-thalassemia gene detection has the advantages of being high in amplification efficiency, high in probe and hybridization template combination efficiency, higher in detection sensitivity and better in specificity and accuracy.

The invention provides a primer combination and method for detecting alpha-thalassemia gene mutation based on a high throughput sequencing technology. The primer combination comprises a specific amplification HBA gene and single nucleotide polymorphism (SNP) primers closely linked within the 2Mb ranges of the upstream and the downstream of the specific amplification HBA gene. The method has the advantages of universality, multi-site SNP sequencing, high throughput, low cost, high sensitivity and strong specificity, all alpha-thalassemia mutation can be detected before embryo transplantation, the detection process is rapider, and detection results are more accurate.

The invention discloses a noninvasive antepartum fetal alpha-thalassemia gene mutation detection library building method, a detection method and a kit. In the library building method, a specific connector is connected onto a maternal peripheral blood free DNA (deoxyribonucleic acid) fragment; then, a connector connection product pre-amplification product is divided into two parts; two rounds of specific amplification are performed by respectively and independently using forward primers and reverse primers aiming at target sites; the target sites can be enriched at high specificity; the amplification specificity of the primers can be obviously improved. The forward and reverse primer groups of a plurality of SNP (single nucleotide polymorphism) sites used for calculating the fetal free DNA proportion are respectively used for performing two-round specific amplification; the fetal DNA proportion can be efficiently and accurately calculated. The library is subjected to sequence testing; the alpha-thalassemia gene mutation can be accurately and effectively detected; the result is identical to the result of the amniocentesis detection and typing; the safety, the noninvasion and the high efficiency are obviously superior to those of the amniocentesis detection.

The invention relates to a fluorescence quantitative polymerase chain reaction (PCR) detection kit for alpha-thalassemia. The kit comprises probes for detecting SEA, 4.2 and 3.7 type deletion, a forward primer and a reverse primer for amplifying SEA, 4.2 and 3.7 type deletion, a PCR buffer and an enzyme mixed liquor. The kit is convenient and simple to operate, high in sensitivity, and good in specificity, repeatability and stability, and can prevent pollution of a PCR product. The fluorescence quantitative PCR detection of the alpha-thalassemia is carried out by using the kit, the deletion type of the alpha-thalassemia in a peripheral blood sample can be accurately determined, and a reliable experimental evidence is provided for sensitive and early diagnosis of the alpha-thalassemia deletion type.

Pancreatic Neuroendocrine Tumors (PanNETs) are a rare but clinically important form of pancreatic neoplasia. To explore the genetic basis of PanNETs, we determined the exomic sequences of ten non-familial PanNETs and then screened the most commonly mutated genes in 58 additional PanNETs. Remarkably, the most frequently mutated genes specify proteins implicated in chromatin remodeling: 44% of the tumors had somatic inactivating mutations in MEN-1, which encodes menin, a component of a histone methyltransferase complex; and 43% had mutations in genes encoding either of the two subunits of a transcription / chromatin remodeling complex consisting of DAXX (death-domain associated protein) and ATRX (alpha thalassemia / mental retardation syndrome X-linked). Clinically, mutations in the MEN1 and DAXX / ATRX genes were associated with better prognosis. We also found mutations in genes in the mTOR (mammalian target of rapamycin) pathway in 14% of the tumors, a finding that could potentially be used to stratify patients for treatment with mTOR inhibitors.

The present invention provides a primer set for detecting alpha thalassemia and beta thalassemia, and belongs to the field of molecular biology, wherein the primer set comprises an alpha primer set and a beta primer set. The invention further provides a chip capable of simultaneously detecting alpha thalassemia and beta thalassemia, wherein probes having sequences represented by SEQ ID NO:1-31 areimmobilized on the chip. Based on the primer set and the chip, the present invention further provides a kit capable of simultaneously detecting alpha thalassemia and beta thalassemia. The kit of theinvention has advantages of short detection time, low cost, convenient operation, closed tube operation and pollution reducing, and has great significance in the screening of thalassemia population, the genetic counseling and the prenatal diagnosis.

The invention relates to the field of biological medicine, and in particular relates to a gene detection kit for quickly detecting Thailand type alpha-thalassemia in a clinic sample. The technical scheme of the gene detection kit aims at providing the gene detection kit for the Thailand type alpha-thalassemia, wherein the gene detection kit comprises PCR (polymerase chain reaction) liquid, the PCR reaction liquid comprises a forward primer THAI-F and a reverse primer THAI-R, the primers are the forward 10724-10725 position and the reverse 1219-1220 position of the breaking point position aiming at the Thailand type alpha-thalassemia, and the primers are respectively designed at the forward 5'-end and the reverse 3'-end of the breaking point. According to the kit provided by the invention, the leak detection of the alpha-thalassemia caused by the common blood screening method can be reduced, the birth of the children who suffer from the heavy type alpha-thalassemia can be reduced or avoided, and a condition can be created for the more comprehensive thalassemia screening. The detection kit provided by the invention is convenient to use, high in accuracy, and good in social benefit and economic benefit in the high incidence area of the thalassemia.

The invention relates to a nucleic acid composition and a detection kit for detecting genetic anemia as well as a use method. The nucleic acid composition and the detection kit for detecting the genetic anemia can be used for simultaneously detecting four deletion type alpha thalassemia genes, three non-deletion type alpha thalassemia genes, nineteenth mutation type beta thalassemia genes, geneticanemia types such as sickle-shaped thalassemia gene as well as specific gene mutation types. Compared with the prior similar technologies, the detection kit for the genetic anemia has the characteristics that several mutation detection types for the genetic anemia are increased, such as alphaTHAI deleted thalassemia, sickle-shaped thalassemia and 71 / 72(+T) mutation and -28M(A-C) mutation which are relatively-rare thalassemia types. The detection for the genetic anemia types can provide visual reference and prompt for clinically detecting the genetic anemia, so that the leak detection risk ofclinical genetic anemia can be greatly reduced and the birth rate of severe anemia children is reduced.

The invention relates to a gene chip, an amplification reagent and a kit for detecting alpha-thalassemia, and belongs to a molecular diagnosis technology, wherein 3 non-deletion (alpha<CS>alpha, alpha<QS>alpha and alpha<WS>alpha) alpha-thalassemia and 4 deletion (--<SEA>, --<THAI>, -alpha<4.2> and -alpha<3.7>) alpha-thalassemia can be rapidly and simultaneously detected by combining direct PCR andreverse dot blot (RDB).

The invention relates to a gene chip for detecting alpha-thalassemia, a using method thereof and a kit with the same. The gene chip comprises a solid-phase carrier and an oligonucleotide probe fixed on the solid-phase carrier, wherein the oligonucleotide probe is a nucleotide sequence shown as SEQ ID NO. 1-195. The using method comprises the following steps: (1) preparing a sample DNA fragment; (2) fluorescently labeling the DNA fragment; (3) purifying a gDNA labeled product; (4) hybridizing the gDNA labeled product with the gene chip; and (5) washing the hybridized gene chip, and scanning a chip hybridization signal. The invention also relates to the kit containing the gene chip. The gene chip provided by the invention has easy operation steps, is high in detection specificity and good in stability, and can complete a process from extracting samples to obtaining scanning results within two days, thereby being low in cost and applicable to detection for deletion or repeated mutation in patients with alpha-thalassemia and prenatal diagnosis.

The invention relates to the field of a biological medicine, and particularly relates to a real-time fluorescent PCR (polymerase chain reaction) detection kit for non-deletion type alpha thalassemia. The kit comprises two PCR reagents, namely a PCR reagent I and CPR reagent II, wherein each PCR reagent comprises a primer, a probe, PCR Buffer, dNTPs, MgCl2, DNA (deoxyribonucleic acid) polymerase, UNG (unguentum) enzyme and glycine betaine. By adopting the kit, QS (quality safety) and CS mutant types of the non-deletion type alpha-thalassemia can be simultaneously detected by a single test. The real-time fluorescent PCR detection kit has the characteristics of short detection time, high sensitivity and strong specificity, and is simple to operate; the result can be judged and read after fluorescent PCR is finished; the problem of carrying out complicated operations of hybridization, sequencing and solubility curve analysis on a PCR product in the prior art is overcome.

The present invention discloses a purely natural medicine, which is used for treating thalassemia and is made by extracting effective active ingredients of marine lifen and a plurality of types of medicinal materials. The formula comprises the following weight percent: 10 percent of sea cucumbers, 10 percent of sea mud worms, 10 percent of Pelamis platurus, 7 percent of oyster, 7 percent of spirulina, 7 percent of Angelica, 7 percent of radix astragali, 7 percent of large ginseng, 7 percent of prepared rhizome of rehmannia, 7 percent of fructus broussonetiae, 7 percent of leatherleaf milletia, 7 percent of Psoraleae, and dried human placenta. The purely natural medicine has the effects of restoring the hematopoietic function, enhancing the level of hemoglobin (Hb), improving the clinical symptoms, and raising the quality of life. Moreover, the medicine has significant curative effects on alpha thalassemia and beta thalassemia, and has the capability of improving the Hb level, the clinical symptoms and the quality of life. The total efficiency of curative effects can reach 93.55 percent.