Hybrid multi-step nucleic acid amplification
An isothermal nucleic acid amplification and polynucleotide technology, which is applied in the field of compound multi-step nucleic acid amplification
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Embodiment 1
[0125] Thermal cycling preamplification methods include any suitable thermal cycling method, including PCR, reverse transcriptase PCR (rtPCR), and other PCR methods. Two-step PCR methods include methods with the steps of: incubating at a first temperature (e.g., between about 38-45°C, or about 42°C); at an elevated second temperature (e.g., about 90-105°C); between, or about 98°C); thermal cycling between two temperatures, wherein the two temperatures are a) an elevated second temperature (e.g., between about 90-105°C, or about 98°C) and b) a lower annealing temperature (Tm, e.g., between about 50°C and about 80°C); followed by a lower third temperature (e.g., a temperature between about 65°C and about 75°C) under incubation. Three-step PCR methods include methods with the steps of: incubating at a first temperature (e.g., between about 38-45°C, or about 42°C); at an elevated second temperature (e.g., about 90-105°C); between, or about 98°C); thermal cycling between three te...
Embodiment 2
[0152] In one non-limiting example, the rapid identification of acute and definitive HIV infection using a single test capable of detecting HIV nucleic acid, antibodies, and antigens can benefit public health by helping patients with positive HIV results get the care and services they need as quickly as possible. have a huge impact. In one embodiment, a single test for HIV is provided that is capable of detecting HIV nucleic acid, antibodies and antigens (for both diagnostic and prognostic purposes), is easy to use on processed or unprocessed samples, is capable of qualitative and / or Quantitative application can be performed in 5 hours or less, 4 hours or less, 3 hours or less, 2 hours or less, or 60 minutes or less and is suitable for commercial promotion.
[0153] In one embodiment, the HIV test of the invention tests all of the following from a sample obtained from a subject in a single device or under the housing of a device: HIV-1 RNA, HIV-2 RNA, p24 antigen, HIV -1 anti...
Embodiment 3
[0176] Embodiment 3MRSA detects
[0177] Methicillin-resistant Staphylococcus aureus (MRSA) is a genus of Staphylococcus aureus (S. aureus) that can cause infections in humans and is resistant to β-lactam antibiotics. MRSA infection can be difficult to treat due to its resistance to certain antibiotics.
[0178] Staphylococcus aureus usually becomes methicillin resistant by acquiring the mecA gene. The mecA gene is normally located in the staphylococcal cassette mec (SCCmec), a multigenic, transferable genomic element. There are different types of SCCmec, the known SCCmec types are approximately 21,000-67,000 nucleotides in length in size. Typically, in each type of SCCmec, the mecA gene is surrounded by other genes or elements that are other components of SCCmec. In MRSA bacteria, SCCmec containing the mecA gene is integrated into the S. aureus chromosome.
[0179] In order to identify and control MRSA bacteria, effective reagents and methods for MRSA detection are needed...
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