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Sequencing joint and sequencing analysis system thereof

A technology for sequencing adapters and sequencing, applied in the field of molecular biology, can solve problems such as poor detection timeliness

Pending Publication Date: 2022-03-01
HANGZHOU MATRIDX BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, it takes at least 15 hours from the initial sample to the production of results, and the approximate time-consuming process is as attached Figure 1b shown
The timeliness of detection is poor and urgently needs to be improved

Method used

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  • Sequencing joint and sequencing analysis system thereof
  • Sequencing joint and sequencing analysis system thereof
  • Sequencing joint and sequencing analysis system thereof

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Embodiment 1

[0048] A sequencing linker in this embodiment has a Y-shaped structure with partially complementary pairs. One of the strands sequentially includes from 5' to 3': internal Index sequence, Index1 sequencing primer binding region sequence, Index1 sequence, and P7 sequence combined with the chip probe. From 5' to 3' on the other strand, it includes: the P5 sequence combined with the chip probe, the Read1 sequencing primer binding region sequence, the internal Index sequence and the T base overhang. The internal tag sequence regions of the two chains are completely complementary to each other, and the sequencing primer binding region sequences are partially complementary to each other. Its structure is as attached Figure 5 As shown, three lengths of internal Index sequences are used, the lengths are 6 bp, 7 bp and 8 bp respectively.

[0049] In this embodiment, a total of 48 internal Index sequences are designed. They were divided into 16 groups, and each group had 6 bp, 7 bp ...

Embodiment 2

[0057] In order to evaluate the analytical performance of this system, we analyzed two clinically positive samples with the sequencing analysis system of the present invention. Among them, the clinical result of sample 1 was Legionella pneumophila infection, and the clinical result of sample 2 was Citrobacter krusei infection. The analysis time and test results of the two samples are shown in Table 1 below. The sequence numbers of Legionella pneumophila measured in each analysis cycle are shown in the appendix Figure 8 , the number of sequences measured by Citrobacter krusei in each analysis cycle is shown in the appendix Figure 9 .

[0058] Table 1. Statistics of clinical sample detection and analysis time

[0059]

[0060] The analysis results show that in the first report of the ultra-fast analysis with a sequencing read length of 22bp, the system has been able to detect positive pathogenic bacteria sensitively; as the sequencing progresses, the number of detected p...

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Abstract

The invention relates to the technical field of molecular biology, in particular to nucleic acid sequencing, and particularly relates to a sequencing joint and a sequencing analysis system thereof. Wherein the sequencing joint is of a partially complementary and paired Y-shaped structure, and one chain of the sequencing joint sequentially comprises an internal Index sequence, an Index1 sequencing primer binding region sequence, an Index1 sequence and a P7 sequence bound with a chip probe from 5'to 3 '; and the other chain sequentially comprises a P5 sequence combined with a chip probe, a Read1 sequencing primer combination region sequence, an internal Index sequence and a T base suspension from 5'to 3 '. Internal tag sequence regions of the two chains are completely complementarily paired, and sequencing primer binding region sequences are partially complementarily paired. When the internal Index sequence joint combination which has artificially balanced base proportion and has different lengths is used, sequencing and analysis of a plurality of samples can be realized while higher sequencing quality and flux are ensured, the turnover time is greatly shortened, and the detection time efficiency is improved.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to nucleic acid sequencing, and in particular to a sequencing linker and a sequencing analysis system thereof. Background technique [0002] Since the New England Journal of Medicine published the first clinical application case of metagenomic next-generation sequencing (mNGS) for the diagnosis of leptospirosis in 2014, mNGS has made a lot of progress in the identification of new pathogens and the diagnosis of rare and important pathogens, and it has also been recognized clinically. Application of mNGS in the field of acute and severe infection. Pathogenic mNGS refers to the extraction of nucleic acid from samples of suspected infection sites, and the nucleic acid fragments are attached to DNA adapters that can hybridize with sequencing chips. The adapters contain index sequences (Index) that can distinguish different samples, through high-throughput sequencing The sequen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/11C12Q1/6869G16B20/30
CPCC12N15/11C12Q1/6869G16B20/30C12Q2525/191C12Q2535/122
Inventor 欧阳川王珺周逸文王江浩刘紫丹
Owner HANGZHOU MATRIDX BIOTECH CO LTD
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