Endotoxin Testing in Pleural Effusion

Pleural effusion is a common clinical feature characterized by the accumulation of pathological fluid in the pleural cavity. The pleural cavity is a potential gap between the visceral layer and the parietal pleura. Normal humans have 5 to 15 ml of fluid in the pleural cavity, which acts as a lubricant during breathing movements. 500 to 1,000 ml of liquid is formed and absorbed in the pleural cavity every day. Any The reason is that the pleural cavity produces more fluid or less absorption, which can produce pleural effusion. According to its mechanism, it can be divided into two types: leaky pleural effusion and exudative pleural effusion. The differential diagnosis of pleural effusion is very complicated. Because of the high morbidity and mortality of bacterial pleural effusion, rapid diagnosis of bacterial pleural effusion is of great clinical significance. Traditional biochemical markers lack specificity, while Gram staining is less sensitive. Although pleural effusion culture has high specificity, it takes a long time. Therefore, it is very important to find biochemical markers for rapid diagnosis and differential diagnosis of bacterial pleural effusion, especially for the testing of endotoxin.

Diagram of fluid buildup in the pleura.Fig1. Diagram of fluid buildup in the pleura.

Method

Pleural fluid LPS levels were determined quantitatively using the Chromogenic Tachypleus Amebocyte Lysate (TAL) endpoint assay.

The following briefly describes the steps used to perform the analysis:

l) Bring the stored samples and kit reagents and materials to room temperature (20-25 "C) before starting the measurement.

2) Preparation of standard solution: by diluting 0.1 ml of endotoxin stock solution with TAL reagent water in endotoxin-free test tubes, prepare a solution containing 1.0 EU/ml endotoxin. 0.5 ml of this 1.0 EU/ml solution was transferred to 0.5 ml of TAL reagent water and labeled as 0.5 EU/ml. Then transfer 0.5 mI of a 1.0 EU/ml solution to 1.5 ml of TAL reagent water and label it as 0.25 EU/ml. Finally, take 0.1 ml of 1.0 EU/ml to transfer the solution to 0.9 ml of TAL. Reagent water, labeled 0.1 EU / ml. TAL reagent water was used as a negative control.

3) 100μl of each sample or standard was transferred to pyrogen-free tubes and incubated with 100μl of tachypleus amebocyte lysate at 37°C for 8 min.

4) 100μl of the chromogenic substance was then added and incubated at 37"C for 6 minutes.

5) 500μl of each of the three provided azo-reagents was added to the mixture in the tubes.

6) UV absorbance at 545nm was measured.

7) A standard curve was made and the level of endotoxin in each sample was determined.

Our Services for endotoxin testing in pleural effusion

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 pleural effusion
Testing purpose Finding biochemical markers is very important for the rapid diagnosis and differential diagnosis of bacterial pleural effusion, especially for the testing of endotoxin.
Testing cycle 3-5 days.
Service including We provide you with raw data and test reports.
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References

  1. Iwanaga, S. Biochemical principle of Limulus test for detecting bacterial endotoxins. Proceedings of the Japan Academy. Series B, Physical and biological sciences. 2007, 83 (4): 110–9.
  2. Porcel JM; et al. Pleural effusions due to pulmonary embolism. Current Opinion in Pulmonary Medicine. 2008,14 (4): 337–42.

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