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Transcript
Saliva and saliva spot inspection 钟树荣(PhD,副教授) 昆明医科大学法医学院 Cell Phone:18213823198 E-mail:[email protected] What are the objects found saliva stain easily ? bite mark 一、The characteristics of the saliva and saliva stain • (一)、saliva • (saliva) is secretion of the salivary glands, saliva is colorless, tasteless, and is close to neutral liquids. water (99.4%) 无机 物质 Sodium, potassium, chloride, calcium Phosphoric acid Carbonic acid rhodanic acid saliva Blood type thing 有机 物质 Mucin, ball eggs Amino acids, urea, urine 淀粉酶 麦芽糖酶 • Fall off in addition, the saliva and oral mucosa epithelial cells, a large number of bacteria, a small amount of white blood cells, and food scraps, etc (二)、Saliva spot • Saliva stain is saliva drying after the formation of the spot mark, saliva spot is more common in forensic inspection of one of the biological material inspection • Saliva spot on a white background with pale yellow, Pale blue fluorescence under ultraviolet light 二、Saliva and saliva spot inspection purpose 个人识别 (Individual Identification) 亲权鉴定 (parentage testing) Confirm the saliva spot 三、Saliva stain proof Confirm whether or not Saliva spot Salivary amylase Check out the test Fall out of the oral mucosa epithelial cells Check out the test (一)、Saliva amylase detection test Starch iodine turned blue theory iodine starch Flecked leaching liquid The same blue Change the blue 氯化三苯基 四氮唑 In the spot amylase 碱性铜 Prove the existence of sugar In the spot amylase matters need attention Flecked unchanged leaching liquid reaction with starch and iodine blue, can only prove flecked contains amylase, doesn't mean the blackspot is saliva. (二)、Fall off oral mucosal epithelial cells of the inspection cell membrane Flat, polygons, the shape is not very regular Cytoplasm nucleus HEDyeing Acid fuchsin methylene blue staining Cell membranes are thin, coloring shallow in the cell membrane is the cytoplasm, dyed deep shape larger is the nucleus. attention: Because saliva spot check material often appear in People's Daily supplies Or items, such as cups, cigarette butts, stone, etc., so confirmed Saliva after the spot, can be directly used to identify the individual, don't have to Species identification. But when determining the bite of saliva spot, first of all want to make sure Is a person or animal bites. 四、Saliva of personal identification Determination of saliva secretion spot ABO blood group and the state Polymorphisms of proteins and enzymes in saliva spot detection Sex identification of saliva Saliva of DNA polymorphism analysis (individual identification, paternity test) Carrier Type DNArelative amount About the data Fresh saliva many 0.8ug/ml Sterile physiological salt Water gargle water more 1.5ug/ml Saliva swabs few 0.26ug/cm2 Saliva spot less 0.16ug/cm2 cigarette end less 12-120ng/个 The PCR technology can enlarge the existing DNA is extremely limited to 2 ng, Reason for common saliva (spot) for examination for DNA testing is absolutely Practical and feasible. The extraction of DNA specimens 1. Biological samples and the separation of biological components (carrier) 2. DNA extracted from cells and cells components 概述 with other How to obtain nuclear DNA ? summarize Raw material cells SDS hypotension Nuclear DNA extraction principle nucleus SDS Heated to boil Nuclear membrane rupture, released from within the nuclear DNA Proteinase Hydrolysis of DNA binding proteins, DNA free in solution Magnetic beads, silica gel Phenol and chloroform, ethanol Impurity removal, collecting DNA 概述 The ideal DNA extraction (1)Maximum remove the RNA in the sample DNA solution, the composition such as proteins, polysaccharides and lipids molecules; (2)As far as possible keep the integrity of the extracted DNA molecules; (3) to reduce or eliminate the possible of enzyme inhibition substances, such as organic solvents, detergents, metal ions, and dyes, and other enzyme activity by inhibiting DNA analysis. summarize 五、Saliva and saliva of extraction (1)、Fresh saliva collection : First gargle, gauze or cotton swabs to be included in the oral cavity, take out after soaking, dry in ventilation places immediately cool, package equipment inspection, respectively. If gathering flow of saliva, because it contains blood enzymes, shall be immediately put in boiling water bath (bath boil 10 min) inactivated enzyme type material. (2). Saliva (spot) extract on the site Found at the scene of the cigarette butts, paper towels, cups, etc should be wearing gloves or with tweezers collection, packaging, respectively. The scene suspected phlegm article such as stains, skin and bite marks, soaked with saline water gauze wipe patches, transferred to dry patches on the gauze shade, mark Second step transfer method • Key to the success of saliva spot transfer : Drink or jelly contact people in the sales process, the scene of the mug is not only the criminals used, large area extraction line won't work for sure. This requires inspection case personnel should through the simulation experiment in extraction for examination or normal life habit that target area. Several kinds of common carrier transfer of saliva on the spot With a handle mug (1)pouring cup Cup without handle (2)Mineral water bottle (3)butt (4)Jelly shell (5)bite mark Generally associated with traces of saliva when occlusion, so it cannot be ignored Wound saliva spot inspection. Found after a bite, use dry first Net gauze to cover, to prevent pollution. Then wipe with two step method Pass the effective components in saliva stain. Do not use hand before inspection Wipe, also don't blot out drugs. (6)Sunflower seeds Due to the different people's habits, into hard seeds with shell on the adhesion of the mouth How much cavity epithelium also will be different. But according to the habits of most people , is to use teeth clacking, so it contains relatively few components . Sunflower seeds in the fiber and has strong adsorption, should not be so Should use the method of transfer cells, direct access to the sunflower seeds. In addition, Sunflower seeds on the surface due to contain pigments, add baking in the process of making And have serious effects on amplification so using chelex extracted to The DNA to do further purification and enrichment. (7)Gum or coffee candy Summary Saliva and saliva spot inspection: • perusal • Saliva stain proof (amylase, shedding of the oral mucosa epithelial cells) • DNA polymorphism analysis (individual identification and paternity tests) • draw