Endotoxin Testing in Bronchoalveolar Lavage Fluid

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Gram-negative pneumonia is one of the most common nosocomial pneumonias, with a mortality rate of up to 50%. Prompt diagnosis and effective use of antibiotic treatment can reduce the mortality rate, so it has important clinical significance. Prospective trial to detect bronchoalveolar lavage fluid (BALF) endotoxin using the radon cell lysate test from January 1997 to assist rapid diagnosis of Gram-negative pneumonia.

Bronchoalveolar lavage fluidFigure 1. Bronchoalveolar lavage fluid.

Testing method

The clinical diagnosis of pneumonia is based on the appearance of new or progressive lung infiltrates on the chest radiograph with one of the following conditions: ① purulent bronchial secretions; ② body temperature above 38.5°C or below 36.5°C; More than 10×109/L; ④ Radiological manifestations of lung abscess formation. For bronchoscopy, Olympus BF-30 fiberoptic bronchoscope was used, as usual. Bronchoalveolar lavage (BAL) was performed with fiberoptic bronchoscope catheter method, that is, after the fiber bronchoscope reached the area of ​​interest, it was inserted through the fiberbronchoscope working tube With a thin catheter, 0.9% sodium chloride 50 mL was injected into the bronchial subsegment or subsegment through the catheter 2 times in total, and the syringe was connected for direct negative pressure suction. The first rinse was used for culture, the second rinse was used for LAL test, cell count and intracellular bacteria examination. Bacteriological examination was performed within 1 hour after specimen collection. Under sterile conditions, 50 μL of bronchoalveolar lavage fluid (BALF) was dripped onto sheep blood plates, and spread with a seed ring. After the surface is dried and cultured, the colony count is multiplied by the dilution factor to obtain the number of colonies per milliliter (cfu), and the number of colonies per liter of cultured colonies reaches 106 cfu/L or more is the basis of bacteriological diagnosis of pneumonia.    Limulus Amebocyte Lysate test uses semi-quantitative method, and intracellular bacteria are tested by Gram stain.

LAL test is a kind of arthropod. Amebocytes in the blood contain factor C, factor B and proclottin-gEnzyme, which can be activated by endotoxin. The activated coagulase can make the coagulation in the deformed cells coagulagen is coagulated to form coagulated proteins, which can be used for qualitative, semi-quantitative and quantitative determination of endotoxin. The reaction has high sensitivity (pg/ml) and specificity. It can be divided into gel method, turbidity method and color matrix method.

Our Services for endotoxin testing in bronchoalveolar lavage fluid

Creative BioMart offers a corresponding endotoxin testing service. You can purchase the corresponding endotoxin removal kit and related accessory products according to the needs of your own samples. We guarantee that all instruments, water, reagents, and consumables used in the experiment are free of endotoxins, and the experiment is conducted in a clean room to ensure that low levels of endotoxins are returned to the sample. In addition, we can also provide you with related services including:

Our solutions

Project name Endotoxin testing in bronchoalveolar lavage fluid
Testing purpose Testing of endotoxin in bronchoalveolar lavage fluid (BALF) to help quickly diagnose Gram-negative pneumonia, timely diagnosis and effective use of antibiotic treatment can reduce mortality.
Testing cycle 3-5 days.
Service including We provide you with raw data and test reports.
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Conclusions

LAL is sensitive to endotoxin, with simple method and high accuracy. The report can be obtained within 1 hour after the test. It has been widely used in the examination of serum, cerebrospinal fluid and other body fluids. When using BAL to diagnose pneumonia, if it is listed as a routine examination, it will undoubtedly improve the speed and accuracy of diagnosis. If a new purine reagent with higher purity can be used, false-positive reactions caused by β-D-glucan can be avoided, thereby further improving the accuracy of the test

Reference

  1. Baker AM; et al. Decision making in nosocomial pneuonia: an analytic approach to the interpretaion of quantitative bronchoscopic culture. Chest, 1995, 107 (1): 85-95.

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