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A method for identifying the tissue origin of microfluid plaques

A body fluid and trace technology, applied in the field of forensics, can solve the problems of loss of RNA template, limited genetic material content, and inapplicability

Active Publication Date: 2020-03-06
SICHUAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this extraction method needs to be based on a certain amount of body fluid, and a large amount of RNA template will be lost for traces of body fluid, so it is not suitable for traces of body fluid.
The total amount of genetic material available for analysis in micro-fluid spots is often very limited, and it is difficult to obtain ideal results using traditional reverse transcription detection methods

Method used

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  • A method for identifying the tissue origin of microfluid plaques
  • A method for identifying the tissue origin of microfluid plaques
  • A method for identifying the tissue origin of microfluid plaques

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0128] (Identification of 1, 2, and 4 fingerprints) (1 means that 1 fingerprint was collected in total, 2 means that 2 fingerprints were collected in total, and 2 fingerprints were wiped with the same FTA card; 4 means that 4 fingerprints were collected, and the same FTA card was wiped 2 fingerprints)

[0129] (1) Preparation and collection of body fluid spots

[0130] Fingerprint preparation: Volunteers were not allowed to wash their hands within 1 hour before fingerprint pressing, and the thumb pressed on the slide for 2 minutes.

[0131] Use tweezers to tear off a 3×3mm 1 / 3 thickness FTA card, wet it with 0.1% Triton X-100 and wipe it.

[0132] (2) For each group of samples, add 0.4μl Biotinylated Oligo-dT30VN (100μM), 4μl dNTP (10mM), 0.3μl RNaseOUT (40U / μl), 0.444μlnuclease-free water, 7μl lysate, and mix well at a preheated constant temperature Incubate at 72°C for 3 minutes on the instrument, and immediately place it on ice for 2 minutes. During this period, configur...

Embodiment 2

[0146]Preparation of saliva spots: Take fresh saliva with EP tube, centrifuge at 12000r / 10min, take 5μl of supernatant and drop it on a glass slide, and let it dry overnight at room temperature to simulate trace spots. Other conditions are with embodiment 1.

[0147] result Figure 5 As a result of saliva plaques, a saliva-specific locus, HTN3, was detected.

Embodiment 3

[0149] Semen spot preparation: take fresh semen with EP tube, centrifuge at 12000r / 10min, take 5μl of the supernatant and drop it on a glass slide, and let it dry overnight at room temperature to simulate trace spots. Other conditions are with embodiment 1.

[0150] result Figure 6 As a result of sperm spots, two semen-specific loci were detected.

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Abstract

The invention provides a method for identifying the tissue source of trace body fluid spots, which belongs to the technical field of forensic medicine. FTA cards are used to collect trace body fluid spots; the body fluid spots are cracked and directly reverse-transcribed to obtain full-length cDNA, and the cDNA is subjected to Pre-amplification to obtain a pre-amplification product; purify the pre-amplification product, and perform PCR amplification on the obtained purified product using a primer combination for a specific site, and according to the expression of the specific site in the amplification product, Determine the tissue source of the fluid spot. In the present invention, after using the FTA card to collect trace body fluid spots, all samples are directly lysed and reverse-transcribed to minimize the loss of cDNA, and combined with other methods, the identification of the tissue source of trace body fluid spots is realized, and the identification of trace body fluid spots The sensitivity reaches 1pg.

Description

technical field [0001] The invention belongs to the technical field of forensic medicine, and in particular relates to a method for identifying the tissue source of trace body fluid spots. Background technique [0002] Bodily fluid spots are common forensic biological samples at crime scenes, and determining their histological origin can provide key clues to the nature of a case, such as a sexual assault case. However, with the improvement of the anti-investigation ability of criminals, the obvious biological evidence on the scene is often destroyed consciously, and more of the leftovers are neglected trace or even trace samples. Therefore, how to improve the identification of trace body fluid spots is an urgent need to be solved. problem. [0003] As a more mature body fluid-specific marker in forensic medicine research in recent years, mRNA has focused more on the improvement of specificity and composite detection ability, but there is little research on the detection of ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6888C12Q1/686
CPCC12Q1/686C12Q1/6888C12Q2600/158C12Q2600/16C12Q2537/143
Inventor 张霁王玉芳王秋月陈晓刚高泽华罗海玻
Owner SICHUAN UNIV