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Detection kit of group B streptococcus rapid serological typing and preparation method thereof

A detection kit and serotyping technology, applied in the field of microbial detection, can solve the problems of complex operation, low positive rate and high requirements, and achieve the effects of simple operation, wide application range and high sensitivity

Pending Publication Date: 2019-10-25
厦门市妇幼保健院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The traditional method of separating a single colony on a blood plate for bacterial culture identification is more complicated to operate, has many human interference factors, and has a low positive rate ([4] Shi Chunyan, Qu Shouhui, Yang Lei, etc., Group B Streptococcus carrier status of pregnant women in late pregnancy The impact of detection and carrier on pregnancy outcomes[J]. Chinese Journal of Obstetrics and Gynecology, 2010, 45(1) 12-16); in molecular biology detection methods, PCR detection technology is commonly used, which has strong specificity and high sensitivity , can be operated in batches. Compared with the conventional microbial culture method, the positive detection rate is greatly improved. However, the PCR method has high requirements for the laboratory, requiring a proprietary PCR laboratory and special experimental equipment. It is impossible to obtain GBS strains and conduct drug sensitivity tests. Other shortcomings ([5] Jia Zhonglan, Lu Xin, Xu Lifeng. Effect evaluation of three methods for screening group B streptococcus in pregnant women [J] Marker Immunoassay and Clinic, 2017, 24(3): 338-339)
Compared with the previous two methods, immunochromatography is simple to operate and takes a short time to produce results. In China, colloidal gold immunochromatography is used for rapid antigen detection of GBS, but there is currently no method for directly detecting its serotype, and for The clinical detection of GBS can quickly identify its serotype, which is more meaningful for preventive medicine

Method used

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  • Detection kit of group B streptococcus rapid serological typing and preparation method thereof
  • Detection kit of group B streptococcus rapid serological typing and preparation method thereof

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preparation example Construction

[0048] The preparation method of group B streptococcus detection kit

[0049] (1) Fluorescent particle pretreatment

[0050] Take out the fluorescent particles, mix them with ultrasound, then take out N-hydroxysuccinimide (NHS for short) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC for short), and equilibrate at room temperature 10-30min.

[0051] Take 100 μl of fluorescent particles + 900 μl of 2-morpholineethanesulfonic acid (MES for short) solution with a pH of 5.0, vortex and mix well, then centrifuge for 15 minutes, discard the supernatant, then add 1ml of MES solution, ultrasonically mix and centrifuge for 15 minutes, discard the supernatant clear;

[0052] (2) Activated fluorescent microspheres

[0053] Add 600-800 μl of MES solution, 100-200 μl of 10 mg / mL NHS solution and 10-100 μl of 10 mg / mL LEDC solution, mix well with ultrasound, activate on a rotary mixer at room temperature for 10-30 minutes, then centrifuge for 15 minutes and discard the supernatan...

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Abstract

A detection kit of group B streptococcus rapid serological typing and a preparation method thereof relate to microbiological detection. The detection kit of group B streptococcus rapid serological typing is provided with a kit body and test paper arranged in the kit body. The test paper is provided with a bottom lining, a sample pad, a marking pad, a coating film, and filter paper, wherein the sample pad, the marking pad, the coating film, and the filter paper are successively connected from the left to the right and are arranged on the bottom lining. The method comprises 1) preprocessing fluorescent particles; 2) activating fluorescent microspheres; 3) combining group B streptococcal antibodies; 4) preparing the marking pad; 5) preparing coating antibodies; and 6) assembling strips. The kit and the method are convenient and fast. When the kit and the method are applied to sample detection, people only needs to wait 10 to 15 minutes after the sample is added and then a detection resultcan be acquired. Operation is simple. The kit and the method can be used for group B streptococcus pure bacteria colonies of clinical culture identification, an enriched culture solution and detection and typing of secretion specimens for clinical routine sampling.

Description

technical field [0001] The invention relates to microorganism detection, in particular to a detection kit for rapid serological typing of group B streptococci and a preparation method thereof. Background technique [0002] Group B Streptococcus (group B streptococcal, GBS), also known as Streptococcus agalactiae, is an opportunistic pathogen that resides in the urogenital tract and lower gastrointestinal tract. It has been reported in the literature that pregnant women infected with GBS may cause adverse pregnancy outcomes such as premature delivery, premature rupture of membranes, intra-amniotic infection, and puerperal infection ([1] Landwehr-Kenzel, S. and P. Henneke, Interaction of Streptococcus agalactiae and Cellular Innate Immunity in Colonization and Disease. Front Immunol, 2014.5: p.519); GBS infection in newborns can cause neonatal pneumonia, sepsis, meningitis, toxic shock, and severe cases may be at risk of death ([2] Nielsen, M ., N. Sheikh, E. Fitzgerald, et a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56944
Inventor 叶辉铭陈小丽车宁郑丽娟苏志英郭清顺
Owner 厦门市妇幼保健院
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