Coagulometrica (Clauss): in prezenta unui exces de trombina, timpul de coagulare al unei plasme citratate, diluate (1/10), saraca in trombocite

este invers proportional cu concentratia de fibrinogen. Testul determina nivelul functional de fibrinogen (activitatea).

In the 1700s, the true beginning of the process of detecting clot formation involved the human eye observing how long it would take for blood to coagulate after being removed from an animal or human being. By the 1800s, scientists could examine blood under a microscope to look for increasing turbidity and determine clotting times from this observation.

Numerous advancements took place from 1822 to 1921, including controlling the temperatures during clot formation, passing objects through the blood to detect resistance and the use of capillary tubes to view clot formation. During the 1900s, the use of test tubes escalated in use and this technique (the precursor to the tilt tube methods) has remained through the years a reliable technique to evaluate clotting times.1All of these methods involved the use of whole blood.

Today, coagulation tests involving plasma have almost completely replaced the whole blood tests. Fully automated coagulation analyzers can trace their roots back to the first tests completed by Gram in 1920 using a plasma clotting time with citrate as an anticoagulant, calcium chloride in the reaction, and a temperature of 37 C. These tests were the foundations of the most commonly used coagulation tests-the PT and aPTT. In the past century, numerous technologies have been developed such as the hook and loop methodologies, the Fibrometer, optical clot detection devices and analyzers that utilize steel balls to detect clot formation.

Detection of Clot Formation

In general, coagulation analyzers use one of two approaches to detect clot formation: They either "look" for the clot (optical, nephelometric) or they "feel" for the clot (mechanical, viscosity based).

Optical Clot Detection (Turbidimetry)

The basic premise of optical clot detection is that as the coagulation process occurs inside the cuvette, the optical opacity increases, thereby decreasing the amount of light transmitted through the plasma. Because the change in transmittance (between the initial reading and the final reading) is used to calculate the results, the effect of adverse sample conditions (such as lipemic or icteric samples) can be minimized.

Some optical clot detection systems have the capability to use multiple wavelengths to try to eliminate the issues associated with these visual interferences within a sample. Although different analyzers utilize slightly different versions, Fig. 1 represents a typical optical clot detection system.

Nephelometric Clot Detection

Nephelometry is the measurement of the amount of light scattered at a variety of angles after passing through a particulate solution. An instrument can utilize nephelometric clot detection by passing each optically clear cuvette through a fixed beam of light on a consistent revolution. For clot testing, a solid-state detector measures the light scatter from the fibrin strands as they form. The final clotting curve is based on consecutive readings per reaction, defining the entire clot curve to its completion.

Although different analyzers may utilize slightly different versions, Fig. 2 represents a typical nephelometric clot detection system.

Mechanical Clot Detection

Mechanical measurement methods detect the physical formation of fibrin strands. These fibrin strands inherently attach to a moving mechanical device, which ultimately either completes or opens an electrical circuit. Any test that has fibrin formation as its endpoint can be determined with this method.

The latest technologies utilize small steel balls and a magnetic sensor below the cuvette to detect clot formation. The use of this method eliminates the issues associated with interferences from lipemic and icteric samples and allows for 1/4 volume patient sample and reagent usage.

Although different analyzers may utilize slightly different versions, Fig. 3 represents a typical mechanical clot detection system.

Viscosity-based Detection

Viscosity-based detection systems involve a cuvette with a steel ball on the inside. An electromagnetic field is applied to each side of the cuvette, which creates a constant pendulum swing of the ball. As the plasma begins to clot, the viscosity of the plasma increases, which will decrease the swinging motion of the ball. The variation in the amplitude of this swinging motion is used to determine the clotting time. This method decreases the interferences found with lipemic and icteric samples.

Although different analyzers may utilize slightly different versions, Fig. 4 represents a typical viscosity-based detection system.

Special Coagulation Testing Needs

In addition to routine testing needs, coagulation laboratories continue to add special coagulation assays to their testing menus. This is being driven by a general increase in the number of patients needing these tests and the ability of the vendors to supply testing kits and analyzers at a lower cost than the cost of sending the test out (Tables 1 and 2). The coagulation analyzers produced today must be able to perform chromogenic and immunoturbidimetric assays so that the laboratories have the capability to perform all coagulation tests available on the market. A new offering from Instrumentation Laboratories/Beckman Coulter is the ACL Acustar, which offers a select number of special coagulation assays using chemiluminescent capability (Table 3).

New to the Market

Two instruments have gained FDA clearance for sale in the U.S. in the past yearthe ACL AcuStar for IL/Beckman Coulter and the Destiny MAX from Tcoag US Inc.

The ACL AcuStarallows the hemostasis lab to utilize an advanced testing platform (chemiluminescent technology) and has been designed to fit the needs of the special coagulation laboratory. The ACL AcuStaris easy to use and can automate the manual steps associated with most complex assays. Its test menu will include assays for antiphospholipid syndrome; ACA (IgG and IgM), B2GPI (IgG and IgM), antibodies for heparin-induced thrombocytopenia (HIT) (Total and IgG), and will eventually include assays for von Willebrand Disease (vWF:Rco and vWF:Ag). The instrument can complete these assay results within an hour.

The Destiny MAXplatform is the latest technology available to the high-volume laboratory market and is ideally suited for both routine and special coagulation laboratories. The Destiny MAXis capable of performing mechanical, optical, chromogenic and immunoturbidimetric assays. The analyzer offers a unique closed tube sampling technique that includes verification of the volumes that have been pipetted, the unique ability to calibrate the PT and APTT (using TriniVeriCAL, and uses half the amount of reagent and patient sample to perform the PT, PTT and FIB assay.

What's Next?

In addition to the instruments discussed in this article, others are pending FDA clearances that will only add to the highly diverse and extensive options a coagulation laboratory has when seeking new technologies.

The providers of automated coagulation analyzers will need to keep pace by introducing new technologies that can meet the needs of all coagulation laboratories, regardless of their size. There is an increased need to supply a family of instrumentation that can create true standardization for a group of regional or national laboratories or a group of instruments in a single laboratory. In high-volume laboratories, many facilities look for their coagulation analyzers to be linked to the various laboratory automated systems available.