Alpha-fetoprotein Assay Research Articles

Ratiometric electrochemical immunoassay based on 2D Co/Fe MOF decorated with toluidine blue and Fc-labeled Schiff base for accurate assay of alpha-fetoprotein in clinical serum

The high level of alpha-fetoprotein (AFP) expression is closely related to hepatocellular carcinoma (HCC). Herein, a dual signal ratiometric electrochemical immunosensor based on chitosan-ferrocenecarboxaldehyde-spindle gold (Chit-Fc-SAu) and Co/Fe metal-organic framework-toluidine blue/polydopamine (Co/Fe MOF-TB/PDA) was proposed for quantitative analysis of AFP. Specifically, Chit-Fc-SAu worked as a substrate to trap more primary antibodies (Ab1) generating the first electrochemical signal from Fc. Thanks to the large specific surface area, the synergistic and electronic effects of Co/Fe MOF nanosheets, and the rich functional groups of PDA, Co/Fe MOF-TB/PDA could load more secondary antibodies (Ab2) and signal molecules (TB) providing another amplified electrochemical signal. In the presence of AFP, Ab1-AFP-Ab2 formed a sandwich structure, and as the AFP concentration increased, the peak current ratio of TB to Fc (ITB/IFc) also increased. The dual signal ratiometric strategy can avoid environmental signal interference and achieve signal self-calibration, thereby improving the accuracy and reproducibility of detection. After a series of exploration, this self-calibrated ratiometric immunosensor exhibited a wide linear range (0.001−200 ng mL−1), a low detection limit (0.34 pg mL−1), and good repeatability. When applied to the assay of clinical serum samples, the detection results of ratiometric sensor were consistent with that of commercial electrochemiluminescence (ECL) immunoassay, significantly superior to that of non-ratiometric sensor. The self-calibrated strategy based on ratiometric sensor helps to improve the accuracy of AFP in clinical diagnosis.

Verification of analytical performance of alpha-fetoprotein assay on the Abbott Alinity ci®: Experience of the central laboratory of Mohammed VI University Hospital of Oujda

The aim of our study was the verification of the analytical performance of alpha-fetoprotein determination on the Abbott CI analyzer utilizing the immuno-chemiluminescence method. The verification process was conducted in the biochemistry laboratory of Mohammed VI University Hospital of Oujda. The working methodology adapted is based on the recommendations of the protocol of the French accreditation committee (COFRAC) accreditation technical guide (GTA) 04, by the evaluation of reproducibility and repeatability. The results obtained by this evaluation were overall satisfactory and have meet the recommended criteria set by supplier and the French society of clinical biology. This study shows that the biochemistry laboratory of Mohammed VI University Hospital of Oujda can deliver an accurate and precise results which can be used for clinical diagnosis and decision making.

A novel photoelectrochemical aptasensor for alpha-fetoprotein assay based on a 3D self-supporting TiO2/CdS/BiOI Z-scheme heterojunction

Alpha-fetoprotein (AFP) is a characteristic marker of liver cancer. This study establishes a PEC aptamer sensor using BiOI/CdS/TiO2 heterojunction material and realizes accurate detection of AFP.

B-379 Analytical Performance of Two Tumor Marker Immunoassays on the Atellica CI 1900 Analyzer

Abstract Background The Atellica® CI 1900 Analyzer is an automated, mid-throughput, integrated chemistry and immunoassay analyzer utilizing both Atellica CH and Atellica IM assays. This study was designed to evaluate the analytical performance of the Atellica IM Carcinoembryonic Antigen (CEA) and Alpha Fetoprotein (AFP) assays* on the Atellica CI 1900 Analyzer. Methods The Atellica CI 1900 CEA and AFP assays use the same reagents and calibrators as the comparable Atellica IM assays. Precision and method comparison (MC) were used as performance indicators for the Atellica CI 1900 Analyzer. Precision studies were performed according to CLSI EP05-A3 using quality control, native, and contrived human serum samples. One aliquot of each sample pool was tested in duplicate in two runs per day ≥2 h apart on each analyzer for ≥20 days. MC studies were performed according to CLSI EP09-A3. Individual native and contrived human serum samples were analyzed using the Atellica IM assays on both the Atellica IM and Atellica CI 1900 Analyzers. Results Representative precision and MC results observed for each assay across indicated sample ranges are listed in the table. Slopes determined by the Deming linear regression model were approximately equal to 1. Conclusion Evaluation of the CEA and AFP assays using the Atellica CI 1900 Analyzer demonstrated good precision and equivalent performance compared to the same assays on the Atellica IM Analyzer. *The products/features mentioned here are not commercially available in all countries. Their future availability cannot be guaranteed.

Longitudinal Evaluation of AFP and CEA External Proficiency Testing Reveals Need for Method Harmonization.

The glycoproteins alpha-fetoprotein (AFP) and carcinoembryonic antigen (CEA) have long been approved as biomarkers for diagnosing and monitoring tumors. International Reference Preparations (IRPs) have been around since 1975. Nevertheless, manufacturer-dependent differences have been reported, indicating a lack of harmonization. This paper analyzes data from 15 external quality assessment (EQA) surveys conducted worldwide between 2018 and 2022. The aim was to gain insight into the longitudinal development of manufacturer-dependent differences for CEA and AFP. In each survey, participating laboratories received two samples with different tumor marker levels. Inter- and intra-assay variability was analyzed and the mean 80% and 90% of the manufacturer collectives were compared to the evaluation criteria of the German Medical Association (RiliBÄK). The median EQA results for CEA revealed manufacturer-dependent differences between the highest and lowest collective of up to 100%; for AFP, the median differences mostly remained below 40%. The coefficients of variation were predominantly low for both markers. We concluded that the current assays for AFP and CEA detection are better harmonized than previously reported. The assays displayed a good robustness; however, a narrowing of the current assessment limits in EQA schemes could further enhance the quality of laboratory testing.

Intramolecular fluorescence resonance energy transfer strategy for accurate detection of AFP-L3% and improved diagnosis of hepatocellular carcinoma

Early and accurate diagnosis of hepatocellular carcinoma (HCC) is of significant importance for improving the survival rate and quality of life for HCC patients. The combined detection of alpha-fetoprotein (AFP) and alpha-fetoprotein-L3 (AFP-L3), namely AFP-L3%, can greatly improve the accuracy of HCC diagnosis compared with AFP detection. Herein, we developed a novel intramolecular fluorescence resonance energy transfer (FRET) strategy for sequential detection of AFP and AFP-specific core fucose to improve the diagnosis accuracy of HCC. Firstly, fluorescence-labeled AFP aptamer (AFP Apt-FAM) was used to specifically recognize all AFP isoforms, and total AFP was quantitatively determined using fluorescence intensity of FAM. Then, 4-((4-(dimethylamino)phenyl)azo)benzoic acid (Dabcyl) labeled lectins (PhoSL-Dabcyl) were used to specifically recognize the core fucose expressed on AFP-L3 that does not bind to other AFP isoforms. The combination of FAM and Dabcyl on the same AFP molecule could generate FRET effect, thereby quenching the fluorescence signal of FAM and quantitatively determining AFP-L3. After that, AFP-L3% was calculated according to the ratio of AFP-L3 to AFP. With this strategy, the concentration of total AFP, AFP-L3 isoform as well as the AFP-L3% were sensitively detected. Detection limits of 0.66 and 0.186 ng/mL were obtained for AFP and AFP-L3 in human serum, respectively. Clinical human serum test results showed that AFP- L3 % test was more accurate than AFP assay to distinguish healthy people, HCC patients and benign liver disease patients. Therefore, the proposed strategy is simple, sensitive and selective, which can improve the accuracy of early diagnosis of HCC, and has good clinical application potential.

A new portable toluidine blue/aptamer complex-on-polyethyleneimine-coated gold nanoparticles-based sensor for label-free electrochemical detection of alpha-fetoprotein.

The quantification of alpha-fetoprotein (AFP) as a potential liver cancer biomarker which is generally found in ultratrace level is of significance in biomedical diagnostics. Therefore, it is challenging to find a strategy to fabricate a highly sensitive electrochemical device towards AFP detection through electrode modification for signal generation and amplification. This work shows the construction of a simple, reliable, highly sensitive, and label-free aptasensor based on polyethyleneimine-coated gold nanoparticles (PEI-AuNPs). A disposable ItalSens screen-printed electrode (SPE) is employed for fabricating the sensor by successive modifying with PEI-AuNPs, aptamer, bovine serum albumin (BSA), and toluidine blue (TB), respectively. The AFP assay is easily performed when the electrode is inserted into a small Sensit/Smart potentiostat connected to a smartphone. The readout signal of the aptasensor derives from the electrochemical response of TB intercalating into the aptamer-modified electrode after binding with the target. The decrease in current response of the proposed sensor is proportional to the AFP concentration due to the restriction of the electron transfer pathway of TB by a number of insulating AFP/aptamer complexes on the electrode surface. PEI-AuNPs improve SPE's reactivity and provide a large surface area for aptamer immobilization whereas aptamer provides selectivity to the target AFP. Consequently, this electrochemical biosensor is highly sensitive and selective for AFP analysis. The developed assay reveals a linear range of detection from 10 to 50000pgmL-1 with R 2 = 0.9977 and provided a limit of detection (LOD) of 9.5pgmL-1 in human serum. With its simplicity and robustness, it is anticipated that this electrochemical-based aptasensor will be a benefit for the clinical diagnosis of liver cancer and further developed for other biomarkers analysis.

An electrochemiluminescence resonance energy transfer biosensor based on Luminol-LDH and CuS@Pt for detection of alpha-fetoprotein

Alpha-fetoprotein (AFP) is the best diagnostic marker for hepatocellular carcinoma (HCC) and plays an important role in the general surveillance of the population. Therefore, the establishment of an ultra-sensitive AFP assay is essential for the early screening and clinical diagnosis of HCC. In this work, we designed a signal-off biosensor for ultra-sensitive detection of AFP based on an electrochemiluminescent resonance energy transfer (ECL-RET) strategy using luminol intercalated layered bimetallic hydroxide (Luminol-LDH) as an ECL donor and Pt nanoparticles-grown on copper sulfide nanospheres (CuS@Pt) as ECL acceptor. The (Au NPs/Luminol-LDH)n multilayer nanomembrane synthesized by our intercalation and layer-by-layer electrostatic assembly process not only effectively immobilizes luminol but also significantly enhances the ECL signal. The CuS@Pt composite has well visible light absorption ability and can burst the light emitted from luminol by ECL-RET. The biosensor showed good linearity in the range from 10−5 ng mL−1 to 100 ng mL−1 and a minimum detection limit of 2.6 fg mL−1. Therefore, the biosensor provides a novel and efficient strategy for the detection of AFP, which is important for the early screening and clinical diagnosis of HCC.

Barhi date (Phoenix dactylifera) extract ameliorates hepatocellular carcinoma in male rats

Hepatocellular carcinoma (HCC) is a malignant tumor with limited treatment options. Given this fact, it may be important to develop new molecular targeted therapies from natural products, especially those which are primary sources of effective anticancer drugs with distinct mechanisms. Moreover, the complementary use of traditional herbs or fruit may increase the possibility of finding curative options for cancer. Here we explore the anticancer effects and possible molecular mechanism of Barhi date extract using an HCC rat model. Thirty two male albino rats were arbitrarily allocated into four groups: a negative control group (NCG); a positive control group (PCG), which received CCl4 (1 ml/kg b.wt./ i.p.) twice a week for three months; a Barhi date extract (400 mg/kg b.wt./day/orally) treatment group (DTG) during the third month of CCl4 administration; and a cisplatin (1.5 mg/kg b.wt./ i.p.) treatment group ( CTG) during the third month of CCl4 administration. After treatment we performed biochemical analyses of all groups to assess relative eukaryotic initiation factor 2 alpha (eIF2α), extracellular signal-regulated kinases (ERKs), protein kinase RNA-like endoplasmic reticulum kinase (PERK), poly (ADP-ribose) polymerase (PARP), and CASPASE 3 protein content, and examined expression of the genes phosphatase and tensin homolog (PTEN) and protein kinase B (AKT). We also performed an immunohistochemistry assay for alpha-fetoprotein (AFP). Our data showed higher PARP and CASPASE3 levels and liver enzymes (alanine aminotransferase [ALT], aspartate aminotransferase [AST], and alkaline phosphatase [ALP]) in the PCG compared to the DTG and the cisplatin treatment group CTG. However, we also found a significant decrease in PTEN in the PCG relative to both the DTG and the CTG. We conclude that the anti-tumor activity of Barhi date extract may be mediated by the inhibition of cell proliferation and apoptosis via the ERK /PARP/caspase3 pathway and the AKT/ PTEN signaling pathways.

A sandwich-like configuration with a signal amplification strategy using a methylene blue/aptamer complex on a heterojunction 2D MoSe2/2D WSe2 electrode: Toward a portable and sensitive electrochemical alpha-fetoprotein immunoassay.

Liver cancer is one of the most common global health problems that features a high mortality rate. Alpha-fetoprotein (AFP) is a potential liver cancer biomarker for the diagnosis of liver cancer. The quantitative detection of AFP at an ultratrace level has important medical significance. Using the reaction of the antibody-antigen pair in an immunosensor enables the sensitive and selective AFP assay. Finding a strategy in signal generation and amplification is challenging to fabricate new sensitive electrochemical immunosensors for AFP detection. This study demonstrates the construction of a simple, reliable, and label-free immunosensor for the detection of AFP on a smart phone. Exfoliated two-dimensional (2D) molybdenum diselenide (MoSe2) and 2D tungsten diselenide (WSe2) were employed to modify the disposable screen-printed carbon electrode (SPCE) to use as the electrochemical platform, which is affixed to a small potentiostat connected to a smart phone. The modified electrode offers antibody immobilization and allows detection of AFP via an immunocomplex forming a sandwich-like configuration with the AFP-corresponding aptamer. A heterojunction 2D MoSe2/2D WSe2 composite improves the SPCE's reactivity and provides a large surface area and good adsorption capacity for the immobilizing antibodies. The signal generation for the immunosensor is from the electrochemical response of methylene blue (MB) intercalating into the aptamer bound on the electrode. The response for the proposed sandwich-like immunosensor is proportional to the AFP concentration (1.0-50,000 pg ml-1). The biosensor has potential for the development of a simple and robust point-of-care diagnostic platform for the clinical diagnosis of liver cancer, achieving a low limit of detection (0.85 pg ml-1), high sensitivity, high selectivity, good stability, and excellent reproducibility.

Diagnostic Value of Multislice Spiral Computed Tomography Combined with Serum AFP, TSGF, and GP73 Assay in the Diagnosis of Primary Liver Cancer.

Objective To explore the diagnostic value of multislice spiral computed tomography (MSCT) scan combined with serum alpha-fetoprotein (AFP), tumor-specific growth factor (TSGF), and Golgi protein73 (GP73) assays in the diagnosis of primary liver cancer (PLC). Methods Totally, 60 patients with PLC admitted to The Second Hospital of Dalian Medical University from January 2019 to January 2020 were included in group A, 60 patients with liver cirrhosis were included in group B, and 60 healthy subjects were included in group C. The serum AFP, TSGF, and GP73 levels were determined, and all participants received MSCT scanning. The diagnostic efficacy of MSCT, assays of serum AFP, TSGF, and GP73, and their combined detection was analyzed. Results Group A had the highest levels of AFP, TSGF, and GP73, followed by group B, and then group C. The sensitivity, specificity, positive predictive value, and negative predictive value of MSCT for PLC were 80.0%,91.7%, 82.8%, and 90.2%, respectively, while those of combined detection of MSCT plus serum AFP, TSGF, and GP73 for PLC were 100.0%, 93.3%, 88.2%, and 100.0%. The combined detection was associated with significantly a higher detection rate of PLC versus stand-alone detection. Conclusion MSCT plus serum AFP, TSGF, and GP73 has a higher detection rate versus stand-alone detection, which shows great potential in the diagnosis of PLC.

A controlled recognizing and releasing glycoprotein based on temperature-responsive phenylboronic microgels for colorimetric analysis of complex samples.

It is challenging to capture glycoprotein from complicated samples with both high specificity and high recovery. Herein, we synthesized temperature-responsive poly (N-isopropylacrylamide-4-vinylbenzeneboronic acid) (PNIPAM-VPBA) microgels having enhanced affinity capability towards glycoprotein, resulting signal amplification in colorimetric analysis subsequently. Through temperature control, the target glycoprotein can be captured and released by the reported microgels unbiasedly. The microgels fabricated were characterized in terms of particle size, phase transition temperature, and toxicity. It was revealed that obvious changes in the particle diameter ranged from 600 to 323nm between 25°C and 55°C, the volume transition temperature was close to human physiological temperature, and low toxicity towards HeLa cells after 24h incubation. Using glucose oxidase as a model target, the microgels were applied as adsorbent materials for preconcentration of glucose oxidase, resulting 22-fold signal enhancement and a limit of detection (LOD) as low as 16.7 U/L. In beer sample analysis, satisfied recoveries of 78.7%-101.6% with the relative standard deviations (RSDs) less than 7.2% was observed. Subsequently, the microgels were combined with enzyme-linked immunosorbent assay (ELISA) for alpha fetoprotein (AFP) determination, by immobilizing horseradish peroxidase (HRP) and anti-AFP antibodies on the surface of microgels. A meaningful low concentration range of 2.5×10-2 to 1.0μg/L was obtained for AFP detection using the microgels based colorimetric method with a LOD of 8.4ng/L. In human serum sample analysis, good accuracy was achieved from method comparing with commercial ELISA kit, satisfied recoveries of 86.1%-96.4% with the RSDs less than 6.0% was observed. The proposed PNIPAM-VPBA microgels have great potential applications in the fields of food and clinical analysis.

Performance evaluation of the Elecsys PIVKA-II and Elecsys AFP assays for hepatocellular carcinoma diagnosis.

Background and AimsProthrombin induced by vitamin K absence‐II (PIVKA‐II) is a serum biomarker linked to hepatocellular carcinoma (HCC), showing superiority to alpha‐fetoprotein (AFP) for early disease detection. We aimed to assess the clinical and analytical performance of the Elecsys® PIVKA‐II immunoassay in diagnosing HCC and evaluate PIVKA‐II's technical performance.MethodsSerum samples from adult cases (i.e. patients with a first‐time HCC diagnosis; n = 168) and disease controls (i.e. patients without HCC with an at‐risk condition; n = 208) were assessed. An AFP cut‐off of 20 ng/mL was used to differentiate between HCC cases and disease controls. Clinical performance of the Elecsys PIVKA‐II assay was compared with that of comparator assays (Lumipulse G PIVKA‐II, μTASWako DCP, ARCHITECT PIVKA‐II) using receiver operating characteristic curve analysis to determine the area under the curve (AUC) values.ResultsThe Elecsys PIVKA‐II assay compared favorably with comparator assays. Using a 28.4 ng/mL cut‐off, the Elecsys PIVKA‐II assay detected HCC with 86.9% sensitivity and 83.7% specificity. Clinical performance of the Elecsys PIVKA‐II assay (AUC: 90.8%) was equivalent to that of comparator assays (AUC: 88.3–89.6%). Relatively high PIVKA‐II concentrations were observed for cholangiocarcinoma and pancreatic cancer with the Elecsys assay in specificity panel analyses, indicating that high PIVKA‐II concentrations should not be used alone in the absence of other clinical data.ConclusionsThe Elecsys PIVKA‐II assay showed good analytical performance under routine laboratory conditions, comparing favorably with comparator assays. These findings support the suitability of the Elecsys PIVKA‐II assay as an aid in HCC diagnosis.

Abnormal ECA-Binding Membrane Glycans and Galactosylated CAT and P4HB in Lesion Tissues as Potential Biomarkers for Hepatocellular Carcinoma Diagnosis

Hepatocellular carcinoma (HCC) is one of the most common types of cancer. Despite decades of research efforts, the search for novel biomarkers is still urgently needed for the diagnosis of HCC and the improvement of clinical outcomes. Previous studies of HCC clinical biomarkers have usually focused on serum and urine samples (e.g., serum Alpha-fetoprotein (AFP). However, cellular membrane proteins in lesion tissues are less used in HCC diagnosis. The abnormal expression of membrane glycoproteins in tumor lesions are considered as potential targets for tumor diagnosis and tumor therapies. Here, a lectin array has been employed to screen and identify abnormal glycopatterns and cellular membrane glycans in HCC lesion tissues compared with adjacent non-tumor tissues. We found that there was significantly less expression of Erythrina cristagalli (ECA) lectin binding (Galβ1-3/β1-4) glycans on the cellular membrane of HCC lesion tissues compared with those of adjacent non-tumor tissues. Immunohistochemistry analysis further showed that ECA-binding ability on the membrane proteins of HCC tissues progressively decreased in different tumor-node-metastasis (TNM) stages (stage I to stage III) as the malignancy of liver cancer increased. Receiver operating curve (ROC) analysis showed ECA-binding ability yielding a sensitivity of 85% and specificity of 75%, and a combination of ECA and AFP has better clinical diagnostic efficiency, yielding a sensitivity of 90% and specificity of 85%, than ECA or AFP assay alone. ECA pull-down followed by mass spectrometry further showed that there was significantly less expression of ECA binding membrane catalase (CAT) and prolyl 4-hydroxylase beta polypeptide (P4HB) in HCC tissues compared with the adjacent non-tumor tissues. The abnormally increased expression of total CAT and P4HB and decreased expression of galactosylated membrane CAT and P4HB in HCC cell lines were correlated with an HCC metastasis status. Our findings suggest that abnormal declined ECA-binding galatosylated membrane glycans and two galactosylated-CAT and P4HB glycoproteins in lesion tissues are potential biomarkers in the diagnosis and/or metastasis prediction for HCC.

Development of active affibody aggregates induced by a self-assembling peptide for high sensitive detection of alpha-fetoprotein

Alpha-fetoprotein (AFP) is one of the most important biomarkers associated with primary liver cancer and the development of accurate and sensitive methods for detecting the serum level of AFP is of great importance in cancer diagnosis. In this study, three self-assembling peptides (SAPs) tagged AFP specific affibody dimer (ZAFP D2)2 recombinant protein, (ZAFP D2)2-ELK16, (ZAFP D2)2-L6KD and (ZAFP D2)2-R18A were successfully produced and characterized. The (ZAFP D2)2-ELK16 affibody aggregates showed higher and more specific binding affinity to AFP compared to (ZAFP D2)2 and other two SAP-tagged (ZAFP D2)2 and in addition displayed enhanced AFP-detection sensitivity and improved stability. A sensitive two-site enzyme-linked immunosorbent assay for AFP using (ZAFP D2)2-ELK16 as the detection reagent was also established. The AFP detection limit and linear range using (ZAFP D2)2-ELK16 based two-site ELISA were 9.7 ng mL−1 and 20–600 ng mL−1, which were significantly improved compared to those of the (ZAFP D2)2 based two-site ELISA for AFP. To detect AFP in serum samples, the average recovery ranged from 95.02% to 110.41% and the relative standard deviation was in the range of 2.45%-3.97%, which meet the standard clinical testing requirements. This proposed strategy provides a high yield, easy purification and green biosynthesis method to prepare SAP-based affibody aggregates for enhanced detection sensitivity by improving the immunoassay signal amplification activity.

Biomarker utility for early diagnosis of hepatocellular carcinoma in Egyptians with viral hepatitis C and B

BackgroundThere is an urgent need to identify novel biomarkers for hepatocellular-carcinoma (HCC) patients who cannot be diagnosed early as in very small lesions and alpha-fetoprotein (AFP)-negative HCC reported in 40% of HCC cases.ObjectiveTo investigate the diagnostic efficacy of AFP, lens culinaris agglutinin-reactive AFP fraction (AFP-L3), des-γ-carboxy prothrombin (DCP), and glypican-3 (GPC3) either alone or in combination and the diagnostic efficacy of the GALAD score [sex (G) and age (A), AFP-L3 (L), AFP (A), and DCP (D)] for early detection of HCC among Egyptian patients with virus-related liver cirrhosis, either hepatitis C or -B viruses.Patients and methodsThis case–control study was conducted on 90 patients that were divided equally into three groups: liver cirrhosis (GI), HCC less than 3 cm (GII), and HCC 3–5 cm (GIII). Assay of AFP, AFP-L3, DCP, and GPC3 was done by enzyme-linked immunosorbent assay. GALAD score was calculated.ResultsThe diagnostic value of AFP, AFP-L3, DCP, GALAD, and GPC3 markers to detect HCC lesions less than 3 cm was assessed by the area under the receiver-operating characteristic curve, which was 0.74, 0.8, 0.75, 0.71, and 0.8, respectively. Sensitivity and specificity were calculated for AFP (86.2%, 93.1%), AFP-L3 (97.2%, 95.4%), DCP (93.4%, 86.2%), GALAD (87.2%, 96.2%), and GPC3 (83.6%, 91.4%). On different combinations, the area under the curve was 0.5 with 100% sensitivity and specificity.ConclusionSerum biomarkers AFP, AFP-L3, DCP, and GPC3 complement each other and demonstrated the best performance for early HCC detection.

An evaluation of the performance of the Point of Care Test iCHROMA™ AFP method: Precision and accuracy

The estimation of serum alpha-fetoprotein (AFP) is useful in the diagnosis and monitoring of primary hepatocellular carcinoma, hepatoblastoma, non-seminomatous testicular germ cell tumours and other germ cell tumours. The iCHROMA™ AFP is a lateral flow chromatography, fluorescence immunoassay (FIA) for the quantitative determination of AFP in serum or plasma. In this study, the Boditech iCHROMA™ AFP assay had a very good precision of 9.8%. It showed a very good correlation with the following 12 methods: Abbott Architect (r2 = 0.9705), BioMerieux VIDAS (r2 = 0.9717), Roche Cobas 6000/8000 (r2 = 0.9738), Siemens Centaur XP/XPT/Classic (r2 = 0.9654), Siemens/DPC/Immulite 2000/2500 (r2 = 0.9673), Siemens/DPC/Immulite 1000 (r2 = 0.9670), Beckman Dxl 600/800 (r2 = 0.9676), Roche Elecsys (r2 = 0.9683), Roche Cobas 4000/e411 (r2 = 0.9688), Roche Modular E170 (r2 = 0.9692), SNIBE Maglumi (r2 = 0.9457) and Ortho Vitros 3600/5600/ECi (r2 = 0.9714). In summary, the iCHROMA™ AFP, a rapid point of care test method, has a within-run precision value of less than 10% and excellent correlations with traditional laboratory methods. There is a consistent overestimation with the iCHROMA™ method, which must be taken into consideration when setting a reference range.

Cerebrospinal Fluid Levels of AFP and hCG: Validation of the Analytical Method and Application in the Diagnosis of Central Nervous System Germ Cell Tumors.

The determination of Human Chorionic Gonadotropin (hCG) and Alpha Fetoprotein (AFP) levels on serum and amniotic fluid plays a fundamental role in the diagnosis and follow-up of specific physiological or pathological conditions (e.g., pregnancy, threat of abortion or germ cell tumors). Recently, the quantification of hCG and AFP in other biological fluids has gained great attention to support the diagnosis, prognosis and follow-up of neoplastic diseases deriving from trophoblastic cells, such as germinomas. Most of the commercial kits for hCG and AFP assays are developed to be used on biological fluids such as serum/plasma and/or urine by manufacturing companies. The aim of this work was to evaluate the suitability of the analytical method certified for the use on serum, and/or amniotic fluid for the quantification of hCG and AFP in cerebrospinal fluid, carrying out an internal validation protocol. The data reported here show that the automated immunochemical method is fit for quantification of hCG and AFP in cerebrospinal fluid (CSF), allowing selective and specific diagnosis of secreting germ cell tumors. This is confirmed by the positive correlation between elevated levels of hCG or AFP and the diagnosis of brain tumors.

Dual-recognition immune-co-chemical ECL-sensor based on Ti,Mg@N-CDs-induced and novel signal-sensing units Poly(DVB-co-PBA)-reported for alpha-fetoprotein detection

In this paper, a high-efficiency electrochemiluminescence (ECL) ternary immune-system was firstly constructed for ultrasensitive assay of alpha-fetoprotein (AFP) based on the Ti, N/Mg co-doped carbon dots (Ti,Mg@N-CDs) as brand-new nanocomposite emitter, Poly(DVB-Co-PBA) as novel AFP-signal reporter and coreaction accelerator. A fluorescent microspheres Poly(DVB-Co-PBA), synthesizing based on 4-vinylphenylboronic acid (4-MPBA) and divinylbenzene (DVB), was used as a signal indicator-probe via the specific carbohydrate-AFP interaction to fabricate a dual-recognition label-free immune-co-chemical biosensor. The as-prepared Ti,Mg@N-CDs exhibited high ECL emission of 3.7-folds compared with CDs. Moreover, the large surface area, promising biocompatibility and nitrogen-rich functional group bearing on the nanohybrid was helpful for the antibody adsorption immobilization of AntiAFP/Ti,Mg@N-CDs/GCE. The successful synthesis and enhancement mechanism of the nanomaterials were also characterized through a serious of techniques including HR-TEM, SEM, EDS, FT-IR, UV–vis, PL and CV. With an immune-co-chemical ECL-assay format, target and signal reporter-probe (Poly(DVB-Co-PBA)) were introduced in sequence via specific reaction. Based on multiple signal amplification and dual recognition strategy, a largely linear detection ranges from 2.25 × 10−6 to 2250 ng mL-1 with a low detection limit (1.55 × 10-7 ng mL-1) were obtained via the resultant ECL ternary (Poly(DVB-Co-PBA)-AFP/AntiAFP/Ti,Mg@N-CDs/GCE) system. This proposed new method was applied to detect human serum samples with satisfactory results and acceptable recovery (96.04 % ∼ 111.22 %), indicating a potential application in clinical monitoring of disease biomarkers.

Clinicopathological Study of Orchidectomy Specimen

The present study was a five years descriptive type of study. In this five years study period June 2012 to May 2017, a total number of 62 surgically removed specimens of testing were received in the Department of Pathology of Krishna Institute of Medical sciences and research centre, Karad for histopathological examination. All orchidectomy specimens received from the surgery department in the period of June 2015 to May 2017. Archival data of cases were studied from June 2012. For prospective cases detail record brief clinical history with age, registration number, biopsy number, presenting signs & symptoms were noted, required serum marker assay for Alpha-fetoprotein, Human chorionic gonadotropin and Lactate dehydrogenase were noted wherever possible. The gross specimens received, they were fixed in 10% neutral buffered formalin/ Bouin‘s solution for overnight fixation. In the present study, the maximum number of cases involving the right side of testis. In this study, Germ cell tumours were the commonest found neoplastic lesion comprising of 16 cases that are 88% of all neoplastic lesions. Most common germ cell tumour in our study was seminoma. In seminoma, the most common age group was found to be in the 3rd and 4th decade. The present study shows one NHL case in 69 years old patient.