Clinical Verification Of H-FABP Diagnosis Kit And The Performance Indexs Of Roche Cobas H232Fast Detection NT-proBNP And CTnT Were Measured And Discussed

BackgroundFatty acid binding proteins (FABPs) is a group of low molecular weight (14KD-15the KD or so) cytoplasmic protein, which is combined with long-chain fatty acid. It is distributed in myocardium, small intestine, liver, adipose tissue, brain, epidermis of mammalian tissue cells. There are nine types of FABPs, which are named after their distribution in tissue and they show significant homology in protein sequence. Each kind of FABPs combine different types of fatty acid, thus FABPs play an important role in uptakeing, ransportation and metabolic regulation of fatty acid. Regulating fatty acid metabolism is the common function of all types of FABPs, but presence and activity of each FABP will be different from others under circumstances of different tissue and condition.Because of tissue specificity in distribution and differeces in structure and function among FABPs, on one hand, fatty acid metabolism is able to develop smoothly in different tissues and conditions, on the other hand, it makes diagnosis of some disease easier. FABP is dissolvable protein with low molecular weight,it can be released rapidly to blood and urina when tissue cells get injured.H-FABP exists in myocardium specifically and accounts for4%-8%of all heart dissolvable proteins.Combined with long chain fatty acid in cardiocyte, H-FABP transports the fatty acid from cytomemtrane to lipined and hydrogened positions,then translocates into energy metabolism system and produces Adenosine Triphosphate (ATP) by oxygenolysis,which offers enegy for myocardium contraction.Myocardial ischemia cause heart cell necrosis, H-FABP myocardial cells from leaking into the blood, and the peripheral blood H-FABP rise. We through the observation, the acute myocardial infarction (AMI), acute myocardial infarction after1～2h rise immediately, and continues to12h, high level and high sensitivity, in24h or so restore and close to normal, and myoglobin (Mb) similar. But it rise abnormaly at the AMI occurred early, more specificity than the Mb and C-reaction protein (CRP) more myocardial and its influence is very small by other organizations. It is expected to become a more ideal diagnosis of early biochemical markers AMI when many domestic and foreign scholars study also found H-FABP and Mb have the basic same AMI diagnosis the timeliness, but in the diagnosis of sensitivity and specificity is superior to the Mb.In order to speed up the H-FABP application in our country and the promotion, we study group in the central health care for the special subject, joint Beijing university life academy of sciences and the Hong Kong university of science and technology to complete the antigen H-FABP cloning and expression of and purification, resistance to H-FABP monoclonal antibodies and the length of polyclonal antibody preparation, H-FABP ELISA test (enzyme linked immunosorbent assay, ELISA) build, and H-FABP in AMI early diagnosis value evaluation. AMI is serious harm of life in patients with clinical emergency, rapid diagnosis, timely decision is to save lives important protection, rescue site testing or to the bed detection (point of care testing, POCT) in such disease diagnosis in show the important value, therefore, the recent and lanzhou biological products research and development, the joint H-FABP qualitative to the bed of fast detection kits (colloidal gold method), and going to apply for production number, according to state food and drug administration the in vitro diagnostic reagents clinical research technique guiding principle ", this kit to clinical experiment.ObjectiveEvaluation developed the H-FABP qualitative to the bed detection kit and the state food and drug administration approved the listed the clinical diagnosis of similar reagent of sex and clinical compliance equivalent.MethodSamples were the plasma or serum of in-patient, outpatient and kindergarden personnel from The305th Hospital of PL A and General Hospital of PL A in2011.240samples were enrolled according to inclusion criteria and exclusion criteria. Test products and control products were detected by the standard of parallel, double-blind and controlled trails. SPSS13.0was used for statistical description and analysis of obtained data. McNamara’s Test was used for paired data and Kappa test was used to see the consistency in the results of two products. P<0.05was considered statistically significant while using two-sided test.Result1.1The contrast product result as standard, the test products and contrast products in A and B two clinical trials in the determination of the agency agreement rate and not consistent rate results show that two clinical test units the measured results of consistency there is no difference (χ2=0.817, v=1, P>0.05), determination of data can be combined analysis.1.2The contrast product result as standard, the sensitivity, specific degree and the total coincidence of test products’results with the comparison were94.92%, 100%and97.5%in clinical A; The sensitivity, specific degree and the total coincidence of test products’results with the comparison were96.43%,100%and98.33%in clinical B; After the merger of two results, the sensitivity, specific degrees and total coincidence of test products’results with the comparison were95.65%,100%and97.92%. The consistency between test products and control products was obvious (Kappa index was0.958). These results show the quality of the test products and comparison product are equivalen.1.3The clinical diagnosis as standard, the positie predictie alue, negatie predictie alue and the total coincidence of test products’ results were100%,98.36%and99.17%in clinical A; the positie predictie alue, negatie predictie alue and the total coincidence of contrast products’results were100%,93.75%and96.67%in clinical A. The positie predictie alue, negatie predictie alue and the total coincidence of test products’results were100%,93.75%and96.67%in clinical B; the positie predictie alue, negatie predictie alue and the total coincidence of contrast products’ results were100%,90.91%and95%in clinical B. After the merger of two results, the test products’positie predictie alue was100%, negatie predictie alue was96%and the total coincidence was97.92%; the contrast products’positie predictie alue was100%, negatie predictie alue was92.31%, and the total coincidence was95.83%. These results show that test products have a good positie predictie alue, negatie predictie alue and clinical compliance.Conclusion1、The quality of the H-FABP detection kit produccted by LanZhou biological research institute and similar listed product produccted by KangBao company production in ShenZhen has the equivalent sex.2、The H-FABP detection kit produccted by LanZhou biological research institute has a good clinical compliance. BackgroundAcute Coronary Syndrome (ACS)(especially AMI) and Heart Failure, which are emergent and severe cardiovascular cases, are threats to health and life of those who take health care. So early and unambiguous dianosis at the scene is significantly important for more hopeful prognosis as well as better life quality.As a marker of myocardiolysis, cardiac troponin (cTn) is one of the important objective criterions for the diagnosis of MI. In October,2007, an experts-allied consensus on "universal definition of myocardial infarction" was formulated and published by a panel of experts from European Society of Cardiolog, American College of Cardiology, American Heart Association and World Heart Federation. According to the new definition, MI diagnosis should be made when both of the following two criterias have satisfied:1) detection of rise and/or fall of cardiac biomarkers(preferably troponin) with at least one value above the99th percentile of the upper reference limit;2) at least one evidence of ischemia, such as symptoms of ischemia, ECG changes of new ischemia(new ST-T changes or new LBBB),development of pathological Q waves in the ECG, imaging evidence of new loss of variable myocardium or new regional wall motion abnormality. Besides, level of cTn can offer a valuable guideline for the treatment (such as acute thrombo lysis and intervention) of ACS.Cardiac troponin (cTn), including cTnI and cTnT,is a subunit of cTn adjustment complex. cTn stems from cytoplasm in the early period of MI while the subsequent continuous cTn is released by actin and myosin from damaged areas. When chest pain occurs, cTnI and cTnT in serum will be detected3-6hours later, and the level of them will keep elevating10-14days after onset of disease. It is generally acknowledged that cTn is of vital importance for myocardial injury diagnosis, and the recently published universal definition of MI further emphasizes great MI dianosis value of cTn.Generally, cTn level is related to infarction size, when elevated, it can not only reflect irreversible myocardical injury, but also plays significant roles in the following aspects:prediction of complication and therapy effect; ACS hierarchy according to risk degree, and guidance for clinical intervention treatment. So, clinical value of cTn has shown in many cases.Brain natriuretic peptide (BNP), a kind of peptid neurohormone, is secreted maily by heart ventricles.Its function is to regulate blood pressure and keep balance of water and electrolytes. Biological active BNP and inactive N-terminal pro-B-type natriuretic peptide (NT-proBNP) will be released from Brain natriuretic peptide precursor (proBNP) when left ventricles fail to function normally. An abundant of recent evidence shows that detection of BNP or its analogs is of great value for diagnosis and prognosis in cardiovascular system disease.What’s more, with the publishment of clinical and biochemical guideline, more and more laboratories are demanded to detect BNP as well as NT-proBNP. Recent study shows that, BNP takes important role in detecting evaluating cardiovascular disease of ordinary people.When togethered with echocardiogram, BNP is especially valuable for diagnosis of acute and chronic cardiac failure. In addition, heart failure patients without any symptom and patients with diastolic dysfunction can be found by noticing fluctuation of BNP level, and dynamic monitor for HF is available because of BNP fluctuation. So, its detecton can improve the accuracy of AHF dianosis compared with estimation only according to clinical symptoms. Negative NT-proBNP, predictive value of which is98%, can help to deplete HF disnosis. A series of evidence prove that NT-proBNP is pretty useful for prognosis evaluation of acute coranaria syndrome, especially acute MI. NT-proBNP is an independent risk factor in high risk group. So further therapy should be taken when NT-proBNP level elevates.For elderly subjects, disease (ACS or HF) tends to show atypical symptoms due to its own chronic diseases of multiple systems. So objective detection are more reliable for early diagnosis of cardiovascular cases, which makes the two markes mentioned above more valuable. With characteristics of old aged and multiple chronic diseases,subjects who need health care are likely to convert their disease to multiple systems failure and even to die if not timely diagnosed and properly treated. Thus, it is undoubtly critical to test the markers promptly in such cases.However, current clinical detection of cTnT and NT-proBNP is almost done in central laboratory of hospital. Severaly hours or even one day is needed for the detection, which severely reduces the guideline value for diagnosis and treatment. Beside detection has been carried out in foreign countries and has been proved more rapid and stable. Nowadays, Emergency and ICU nurses of some medical institutions abroad are trained to do prompt beside detection of the two markers for patients who seem to have heart disease. According to reports, the rapid detection method can significantly shorten the test time compared with traditional methods in central laboratory. Roche Elecsys2010automatic immunoassay analyzer is a world leader in the detection of cTnT and NT-proBNP, and now Roche Company launches Roche cobas h232portable detector, which can rapidly detect cTnT and NT-proBNP. Starting from the actual demand for health care, we validated the clinical effect of Roche cobas h232portable hand-held blood analyzer for rapid detection of cTnT and NT-proBNP. ObjectiveBy evaluating the reliability of Roche cobas h232portable hand-held blood analyzer for rapid detection of cTnT and NT-proBNP, we aimed to confirm the acceptance of rapid detection method,which can improve treatment levels in the performance of severe cardiovascular event for health care subjects.MethodEmergent patients with suspected acute coronary syndrome and AHF patients, both of which come from The305th Hospital and The General Hospital of PLA, were enrolled in our study. Roche cobas h232was used to detect NT-proBNP and cTnT of76clinical samples with whole blood, meanwhile, plasma was saved to detect NT-proBN and cTnT by Elecsys2010.Clinical performance of Roche cobas h232was evaluated by detection of NT-proBNP and cTnT based on precision test, recovery test and anti-jamming test. Software of SPSS13.0and Excell table were used for statistical description and analysis of obtained data. P<0.05is considered statistically significant.Result2.1Correlation experiment:After detecting NT-proBNP by Roche cobas h232and Elecsys2010respectively, correlation coefficient R=0.961within the range of60-9000pg/ml; while in the range of60-3000pg/ml, R=0.948Thus,they showed good correlation(P<0.001).2.2Precision test:For NT-proBNP detection, coefficient of variation within group was2.2%-6.6%, and coefficient of variation between groups was4.6%-5.9%; For cTnT detection, coefficient of variation within group was4.8%-7.4%, and coefficient of variation between groups was5.2%-7.6%.2.3Recovery test:In the use of Roche cobas h232, recovery rate was96.3%-98.6%for NT-proBNP detection and recovery rate was96.7%～101.4%for cTnT detection.2.4Anti-interference test:hemolysis hemoglobin to normal NT-proBNP and cTnT determination value of affected (±15%), for abnormal NT-proBNP and cTnT determination value of less effect (±10%); High concentrations of total bilirubin led directly to the cannot measure NT-proBNP, the determination of the value of normal cTnT affected (±15%), cTnT less influence abnormal determination value (plus or minus10%); Triglyceride of normal NT-proBNP and cTnT determination value of affected (±15%), abnormal NT-proBNP and cTnT determination value of less effect (±0%).Conclusion1、Roche cobas h232portable detector NT-proBNP detection performance index is good and clinical acceptable.2、Roche cobas h232portable detector cTnT detection performance index is good and clinical acceptable.