48 results about "Species level" patented technology

An establishment method of a gene library of fecal flora based on high-throughput gene sequencing employs a ''nested PCR'' method for enrichment amplification on 16S rDNA, so as to reduce the host and food residue genome pollution by the maximum; and V3 and V6 are combined for specific amplification and massive parallel sequencing, so as to obtain the a target gene sequence database of the flora. The library can increase the identification of the bacterial flora from Genus level to Species level. The method can maximally avoid the interference of host cell nucleic acid, and completely and efficiently detect 16SrDNA tag sequence of all bacteria varieties, so as to accurately determine the ecological structure of the intestinal flora.

The present invention relates generally to a method for detecting, enumerating and/or identifying microorganisms in a sample. More particularly, the present invention provides a method for determining total microbial content in a sample by detecting the presence of nucleotide sequences associated with all or part of 16S rDNA or its corresponding 16S rRNA or its homologue, functional equivalent or derivative. The nucleotide sequences of the present invention may be used as an indicator of any microorganism and, hence, represents a universal target sequence which is indicative of total microbial content in a sample. The universal target sequence may also be varied to render same genus or species specific or the universal target used to trap microbial DNA or RNA which may be subsequently analyzed by sequence analysis or genetic probe technology. The universal target sequence is useful inter alia to design as universal primers and probes to amplify any microbial-derived genomic sequence, as a means to detect and enumerate total microorganisms and to identify microorganisms in a sample at the genus or species level. Such uses enable improved methods of enviroprotection, bioremediation, medical diagnosis and industrial microbiology. The present invention further relates to the universal target sequence in isolated form and/or primers or probes capable of hybridizing to same and kits for the detection of total microbial content in a sample.

The invention discloses primers for detecting hybrid trembling poplar molecule and a usage thereof, being specially used in quick molecular detection and identification of trembling poplar, liriodendron tulipifera and hybrid trembling poplar, and belonging to the molecular identification field for new plant variety. A pair of SSR primers, i.e. LT008L/R is designed, and can realize specific amplification of a 190bp product inside the trembling poplar, a 180bp product inside liriodendron tulipifera and a 180bp product, and a 190bp product inside hybrid trembling poplar. The primer adopts a pair of specific SSR primers to carry out once PCR amplification reaction, and carries out electrophoresis by use of 8 percent polyacrylamide gel; moreover, judgment can be carried out directly according to product fragment. The primers can be directly used for the species level identification of hybrid trembling poplar, trembling poplar and liriodendron tulipifera.

The emergence of opportunistic and antifungal resistant strains has given rise to an urgent need for a rapid and accurate method for the detection of fungal pathogens. In this application, we demonstrate the detection of medically important fungal pathogens at the species level. The present method, which is based on a nucleotide hybridization assay, consists of a combination of different sets of fluorescent beads covalently bound to species specific capture probes. Upon hybridization, the beads bearing the target amplicons are classified by their spectral addresses with a 635 nm laser. Quantitation of the hybridized biotinylated amplicon is based on the fluorescent detection with a 532 nm laser. Using this technology we designed and tested various multiplex formats, the performance of forty eight species specific and group specific capture probes designed from sequence analysis in the D1/D2 region of ribosomal DNA, internal transcribed spacer regions (ITS), and intergenic spacer region (IGS). Species-specific biotinylated amplicons (>600 bp) were generated with three sets of primers to yield fragments from the three regions. The developed assay was specific and relatively fast, as it discriminated species differing by one nucleotide and required less than 50 min following amplification to process a 96 well plate with the capability to detect up to 100 species per well. The sensitivity of the assay allowed the detection as low as 10² genome molecules in PCR reactions and 10⁷ to 10⁸ molecules of biotinylated amplification product. This technology provided a rapid means of detection of Trichosporon species and had the flexibility to identify species in a multiplex format by combining different sets of beads. The assay can be expanded to include all known pathogenic fungal species.

The invention relates to the dominance evaluation of ecological communities at the community level and species level, and mainly invents a concept and a method capable of comprehensively evaluating the dominance at the community level and species level. The concept and the method are used to monitor endangered species in macro-ecological communities and to diagnose and predict the risk of human flora-related diseases in microbial communities. The inventor defines a dominance index at the community level, a species dominance distance at the species level and a species dominance index. The threeindexes are defined by a unified mathematical concept and method, and can be used to evaluate the dominance of various biological/ecological communities (including macro-ecological communities (suchas tropical rainforests) and human microbial communities) in various time and/or space scales. In the specification, the microbial flora (community) in the female reproductive tract is taken as an example, and the dominance index is used to study the dominance change of the community in the time scale and the dominance of each species in the community and identify the difference in dominance of the same species in different communities (in the space scale).

Multiplex PCR primers that can amplify the cdt genes of C. jejuni, C. coli, and C. fetus in a bacterial species-specific manner were prepared. Multiplex PCR with the primers was assessed using Campylobacter bacteria, other cdt gene-positive bacteria, and representative bacteria responsible for enteric infection. As a result, the present inventors' multiplex PCR using cdtB amplification primers was proven to enable simultaneous detection of different Campylobacter bacteria with high specificity. The methods of the present invention can identify Campylobacter at the bacterial species level in a single manipulation even when domestic animals or humans are infected with different bacterial species of Campylobacter.

The invention provides a method and device to identify microbial species from a sample. The method includes: acquiring DNA and RNA sequencing results; comparing the DAN sequencing results to obtain afirst candidate microbe set and a specific DNA sequencing read set; comparing the RNA sequencing results to obtain a second candidate microbe set and a specific RNA sequencing read set; using at leastpart of an intersection of the first and second candidate microbe sets as a third candidate microbe set; selecting specific DNA and RAN sequencing reads of same quantity respectively from the specific DNA and RNA sequencing read sets so as to respectively obtain filtered DAN and RAN sequencing read sets; filtering the third candidate microbe set to obtain a fourth candidate microbe set, thereby forming microbial species in the sample. Therefore, the method and device allow species-level microbes to be accurately identified, can avoid interference from background microbes, are suitable for finding actively expressed pathogenic microbes, and are simple to operate.

The invention discloses a library-building sequencing method for detecting full length of bacterial 16S rDNA. By adding a molecular tag UMI to each sample, a ligation library and a splicing library are separately amplified, the two libraries are sequenced, the data is extracted by recognizing the UMI combination, and the full length sequence of 16S rDNA was assembled, and the species type is determined by comparing the databases. The library-building method of the invention is suitable for the detection of the flora structure of all types of samples, and can upgrade the traditional flora structure identification from a genus level to a species level, compared with the genus level, the species level has the advantage that an ecological structure of environmental microorganisms can be more accurately determined, which facilitates deep research, and method realizes the detection of specific bacterial species in samples.

A micro-electromechanical platform and array system and methods for identifying microbial species with single molecule electrical conductance measurements are provided. The electromechanical platform has a two-tier actuation mechanism with a long stroke provided by a comb drive and a fine stroke provided by an in-plane flexural actuator. The platform is capable of making contact with a single-molecule, applying a bias, measuring the current, and performing a large number of measurements for statistical analysis. The system is capable of detecting any microbial species without requiring enzymatic amplification by detecting specific RNA sequences, for example. With oligonucleotide target molecules, the conductance is extremely sensitive to the sequence so even single-nucleotide polymorphisms can be identified. The system can also discern between subspecies using the same DNA probe. The system provides reliable, efficient, and inexpensive detection and species-level identification of microorganisms in complex detecting environments.

The invention discloses a method for evaluating temperature and humidity states of a growing environment of a nursery pig individual based on relative abundance of nasal cavity prokaryotic microorganisms. The method comprises the following steps: quantitatively detecting the relative abundance of the nasal cavity internal door, outline, order, family, genus and species level prokaryotic microorganisms of the nursery pig relative to the nasal cavity total prokaryotic microorganisms, and evaluating the temperature and humidity states of the growing environment of the nursery pig individual by using the relative abundance combination.

The invention discloses a novel detection method of urine bacteria. The bacteria in urine are identified by increasing the amount of culture specimens, changing different growth media and gas conditions, and prolonging the culture time of bacteria. The method comprises the following steps: collection of urine; preservation and transportation; preliminary subculture; sub-subculture; and identification of bacteria. According to the present invention, the kind of bacteria in urine can be determined at the species level, and the further biochemical characteristics and metabolic characteristics ofthe cultured bacteria are analyzed and studied; and missing bacteria by standard culture can be identified by the method, anaerobic bacteria, aerobic bacteria and microaerophilic bacteria that survivein urine can be cultured and identified, and the method of the invention can more fully reflect the situation of all bacteria in urine compared with the standard culture. The time required by the method of the present invention is significantly shorter than the time of 16S rDNA sequencing, and the operation is simpler; the requirements for laboratory and experimental equipment are relatively low,which can be realized by most hospitals and laboratories; and the method has high operability and popularization.

By using an ITS-DNA hybridization technique, microorganisms being quickly differentiated and identified at the bacterial species level. By applying the principle to DNA microarray, environmental microorganisms are detected and identified. By using ITS-DNA hybridization and by applying the ITS-DNA hybridization technique to the DNA microarray, differentiation at the bacterial species level or the relative species level becomes possible.

The invention discloses a gene chip for specific detection of bacillus thuringiensis, and belongs to a soil microorganism gene detection technology. An rpoB unique gene fragment and a 16S unique gene fragment of the bacillus thuringiensis are selected as target sequences for detection according to a reported 16S DNA sequence and rpoB gene sequence of the bacillus thuringiensis, and corresponding multiple PCR amplification primer pairs, probes and detection chips are designed. Accordingly, qualitative and quantitative detection can be conducted on the bacillus thuringiensis in a short time through the gene detection chip, and classification and determination on the bacillus thuringiensis on the species level are achieved; a powerful tool is provided for tracking colonization and reproduction of the bacillus thuringiensis in fertilizer or soil.

The invention discloses a degraded wetland waterfowl diversity recovery method and system. The method comprises the following steps: step 1, collecting existing, peripheral and historical waterfowl species data of a degraded wetland; step 2, according to the three types of waterfowl data, establishing a relationship model, and screening out target species; step 4, according to the target species and the site characteristics, performing partitioning; and step 5, designing and creating each subarea, and applying a strategy in a centralized manner. According to the method, the waterfowl in degraded wetland is innovatively recovered to the original species level before degradation, and degradation of the wetland habitat is restrained, so that waterfowl diversity recovery engineering of the degraded wetland is carried out according to site characteristics on the basis of establishing a historical-surrounding-target species relationship model. The method is clear in target, scientific, simple and convenient, and plays an important role in improving decision-making and engineering efficiency in degraded wetland habitat restoration and waterfowl diversity restoration work.

The invention discloses application of a madder polysaccharide in the preparation of a health-protection food with effects of preventing and treating neurodegenerative diseases. The madder polysaccharide can be a commercially available product or a product prepared by an applicant and is preferably a madder acidic polysaccharide. The invention also discloses a preparation method of the madder acidic polysaccharide. The madder acidic polysaccharide prepared by using the method disclosed by the invention has the characteristics of high polysaccharides content, low impurity content and high uniformity. Furthermore, the preparation method is simple and high in efficiency and is suitable for industrial production. Experiments on an in-vitro aggregation model, mammalian cells and a caenorhabditis elegans disease model show that the madder polysaccharide can effectively suppress the abnormal aggregation of polyglutamine and beta-amyloid proteins and the neurotoxicity of the polyglutamine and the beta-amyloid proteins and also obviously reduce the reactive oxide species level of the disease model, so that the madder polysaccharide has a good preventing and treating effect on the neurodegenerative diseases.

The improved Tm mapping method using imperfect-match linear long quenching probes can accurately distinguish among and identify microorganisms at least at the genus level and often at the species level even in a real-time PCR instrument having measurement errors of Tm values between PCR tubes within ±0.5° C. Therefore, the Tm mapping method can be performed in almost all real-time PCR instruments and can identify unspecified infection-causing pathogenic microorganisms in about 4 hours after sample collection.