35 results about "Reverse Transcription Process" patented technology

The invention provides a viral self-inactivating (SIN) vector on the basis of the avian sarcoma leukosis virus (ASLV) as well as a split-packaging system comprising in addition to the SIN vector a first helper plasmid serving for the expression of the viral fusion protein gag-pol and a second helper plasmid serving for the expression of the retroviral envelope protein (env). The first and second helper plasmid, for example contained in a packaging cell line or transiently transfected, serve for the generation of non-replicating (RCR-incompetent) viral particles containing RNA having a SIN LTR according to the invention at the 3′ terminus, wherein the RNA can have a therapeutically effective section which e.g. is denoted a transgene. This 3′ SIN LTR contains an extensive deletion of the U3 region which in the course of the reverse transcription is copied into the 5′ LTR. In addition, in the SIN vector all coding regions of ASLV as well as the retroviral splice donor site are removed.

The invention provides a positive control composition for a reverse transcription polymerase chain reaction kit and a preparation method thereof, and the positive control composition is characterized in that a non-target virus is taken as positive control, a pair of primers are simultaneously designed for amplification, and the length of an amplified fragment is the same with that of a target gene of the reverse transcription polymerase chain reaction kit; and the non-target virus is inactivated by guanidine isothiocyanate. The positive control prepared by the method is safe, thereby avoiding virus expansion caused by direct use of a virus infection material in the prior art, playing a control role during an RNA (ribonucleic acid) extraction process and an RNA reverse transcription process when serving as the reverse transcription positive control of the RNA virus, proving that an RNA extraction reagent and substances used in a reaction system are normal in activity, facilitating the positive control when serving as the commercial kit and having very high practical value.

The invention discloses a method for positioning and quantitatively detecting 6-methylaminopurine in DNA and RNA. The methylation level of nucleic acid is reflected by detecting 6-methylaminopurine in DNA and RNA, wherein substrate activity in DNA replication and RNA reverse transcription will be reduced due to the existence of 6-methylaminopurine, and in the DNA replication and RNA reverse transcription process, chain extension will not occur when methylation loca occur; therefore, deoxyribonucleoside triphosphate can be doped in for chain extension so as to differentiate the methylation sequence and the non-methylation sequence, and meanwhile the methylation level of a specific locus of DNA and RNA can be quantitatively evaluated. The method can be well used for analyzing 6-methylaminopurine in nucleotide separation of DNA and RNA and can be used for detecting the methylation level of adenine in DNA and RNA easily and conveniently.

The invention relates to a kit for detecting swine fever viruses, which contains RNA (Ribonucleic Acid) enzyme inhibitor, an AMV (Avian Myeloblastosis Virus) reverse transcriptase, Taq enzyme (Thermal-resistant desoxyribonucleic acid polymerase), RNA-free enzyme water, One Step RNA PCR (one-step RNA Polymerase Chain Reaction) mixed solution, a positive and negative reference substance, and mixtures of primers with the sequences shown as SEQ ID NO.1 to SEQ ID NO.6. In the detection process, the reverse transcription process and the PCR amplification process are accomplished in one-step reaction, so that the detection time is shorten to 2-3 hours from the original time of 4-5 hours, thereby benefiting rapid detection. Simultaneously, as the kit uses three pairs of primers, compared with thecommon PCR method, the method has higher sensitivity, thereby contributing to prevention and control of CSFV (Classical Swine Fever Virus) and rejection of recessive poisonous animals, and avoiding resulting in large-range infection. The method is suitable for any laboratories, each-level prevention and control units in the basic government organization, veterinary stations, large, medium and small-size livestock farms and the like.

The invention provides novel high-sensitivity respiratory virus nucleic acid NASBA (nucleic acid sequence based amplification) primers. A respiratory virus conservative area is selected, and a promoter sequence capable of being recognized by T7 RNA (ribose nucleic acid) polymerase is formed at the 5' terminal of one of the synthesized primers. The invention further provides a novel high-sensitivity respiratory virus nucleic acid NASBA detection method. T7 RNA polymerase promoter sequences are added to the 5' terminals of the two primers complementary to two ends of a respiratory virus RNA sequence, so that when the primers enter a circulation phase, RNA sequences [RNA(+)] consistent with a template sequence can be synthesized, RNA sequences [RNA(-)] complementary to a template RNA sequence can be synthesized, and the detection sensitivity of viral nucleic acid is greatly improved; a throat swab sample of a patient can be directly used for amplification, a reverse transcription process is not required, and the operation time is saved.

The invention belongs to the field of research and development of HIV inhibitors, and discloses a preparation method for a terpyridine pyridinium (II) complex and application thereof in HIV reverse transcriptase inhibition. The structure of a cationic moiety of the terpyridine pyridinium (II) complex is as shown in a formula I. A preparation process for the terpyridine pyridinium (II) complex is optimized, the raw material cost is low, and the reaction time is short. The obtained complex is high in purity and yield and has good water solubility and excellent spectral properties. The terpyridine pyridinium (II) complex has the capability of selective binding to a TAR region on HIV RNA, and can block the reverse transcription process of viral RNA by reverse transcriptase and inhibit the replication of viral RNA. The terpyridine pyridinium (II) complex is a highly affinitive HIV RNA selective binding reagent and a highly active HIV reverse transcriptase inhibitor, and is an HIV drug having a great application potential.