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Fulltext Interleukin-6 and intercellular cell adhesion molecule-1 expression remains elevated in revived live endothelial cells following spaceflight

Muid S, Froemming GR, Ali AM, Nawawi H

Malays J Pathol, 2013 Dec;35(2):165-76.
PMID: 24362480 MyJurnal

The effects of spaceflight on cardiovascular health are not necessarily seen immediately after astronauts have returned but can be delayed. It is important to investigate the long term effects of spaceflight on protein and gene expression of inflammation and endothelial activation as a predictor for the development of atherosclerosis and potential cardiovascular problems. The objectives of this study were to investigate the (a) protein and gene expression of inflammation and endothelial activation, (b) expression of nuclear factor kappa B (NFκB), signal transducer and activator of transcription-3 (STAT-3) and endothelial nitric oxide synthase (eNOS) in human umbilical vein endothelial cells (HUVEC) 3 months post-space flight travel compared to ground controls. HUVEC cultured on microcarriers in fluid processing apparatus were flown to the International Space Station (ISS) by the Soyuz TMA-11 rocket. After landing, the cells were detached from microcarriers and recultured in T-25 cm(2) culture flasks (Revived HUVEC). Soluble protein expression of IL-6, TNF-α, ICAM-1, VCAM-1 and e-selectin were measured by ELISA. Gene expression of these markers and in addition NFκB, STAT-3 and eNOS were measured. Spaceflight induced IL-6 and ICAM-1 remain elevated even after 3 months post spaceflight travel and this is mediated via STAT-3 pathway. The downregulation of eNOS expression in revived HUVEC cells suggests a reduced protection of the cells and the surrounding vessels against future insults that may lead to atherosclerosis. It would be crucial to explore preventive measures, in relation to atherosclerosis and its related complications.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Fulltext Seroprevalance of brucellosis among suspected cases in Malaysia

Jama'ayah MZ, Heu JY, Norazah A

Malays J Pathol, 2011 Jun;33(1):31-4.
PMID: 21874749 MyJurnal

Brucellosis is a zoonotic disease which can be transmitted by direct or indirect contact with infected animal or their products. It is an important public health problem but little is known on brucellosis in the Malaysian population. The aim of this study was to determine the presence of Brucella antibodies using commercial Brucella IgG and IgM ELISA kits (Vircell, SL, Barcelona Spain). A total of 184 sera from suspected patients were received from 16 hospitals in Malaysia over the years 2004 to 2009. Only 10 serum samples (5.4%) were positive for Brucella antibodies in which 5 showed the presence of both IgM and IgG. Most of the positive patients were occupationally involved with animals. This study suggests the seroprevalance of brucellosis among individuals who have contact with infected animals in Malaysia is low.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Fulltext A comparative evaluation of dengue diagnostic tests based on single-acute serum samples for laboratory confirmation of acute dengue

Chua KB, Mustafa B, Abdul Wahab AH, Chem YK, Khairul AH, Kumarasamy V, et al.

Malays J Pathol, 2011 Jun;33(1):13-20.
PMID: 21874746

A prospective study was carried out to evaluate the sensitivity of dengue NS1 antigen-capture ELISA in comparison with dengue virus isolation, conventional RT-PCR and real-time RT-PCR for laboratory confirmation of acute dengue based on single-acute serum samples. Four primary healthcare centres were involved to recruit patients with clinical diagnosis of dengue illness. Patient's demographic, epidemiological and clinical information were collected on a standardized data entry form and 5 ml of venous blood was collected upon consent. In the laboratory, six types of laboratory tests were performed on each of the collected acute serum sample. Of the 558 acute serum samples collected from 558 patients with clinical diagnosis of dengue from mid-August 2006 to March 2009, 174 serum samples were tested positive by the dengue NS1 antigen-capture ELISA, 77 by virus isolation, 92 by RT-PCR and 112 by real-time RT-PCR. A total of 190 serum samples were tested positive by either one or a combination of the four methods whereas, only 59 serum samples were tested positive by all four methods. Thus, based on single-acute serum samples, 190 of the 558 patients (34.1%) were laboratory-confirmed acute dengue. The overall test sensitivity was 91.6%, 40.5%, 48.4% and 58.9% for dengue NS1 antigen-capture ELISA, virus isolation, conventional RT-PCR and real-time RT-PCR respectively. Statistically, dengue NS1 antigen-capture ELISA was the most sensitive and virus isolation was the least sensitive test for the laboratory confirmation of acute dengue based on single-acute serum specimens. Real-time RT-PCR was significantly more sensitive than the conventional RT-PCR.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*
Fulltext Urinary type IV collagen levels in diabetes mellitus

Sthaneshwar P, Chan SP

Malays J Pathol, 2010 Jun;32(1):43-7.
PMID: 20614725 MyJurnal

Type IV collagen is the principal component of glomerular basement membrane and messangial matrix. Studies have shown increased levels of urinary type IV collagen (uIV) in diabetic patients compared to healthy controls. The concentration of uIV increases gradually as diabetic nephropathy progresses.
AIM AND METHOD: This study was carried out to determine whether urinary type IV collagen (uIV) can serve as an indicator of diabetic nephropathy. Using a sandwich enzyme immunoassay technique, uIV excretion was determined in 30 type 2 diabetic patients with normoalbuminuria and 20 patients with microalbuminuria.
RESULTS: uIV excretion was significantly increased in type 2 diabetics, in both normoalbuminuric and microalbuminuric patients, compared with healthy controls. The increase in urinary type IV collagen was well correlated with the amount of urinary albumin but not with HbA1C.
CONCLUSION: Our findings that uIV is higher in those with microalbuminuria and correlates with albuminuria, support uIV as an indicator of diabetic nephropathy. Whether the increased uIV excretion would predict the impending renal failure needs further confirmation.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Fulltext Elevated plasma tissue factor levels in neonates with umbilical arterial catheter-associated thrombosis

Tay SP, Cheong SK, Boo NY

Malays J Pathol, 2006 Jun;28(1):41-8.
PMID: 17694958 MyJurnal

Catheterization of the umbilical artery has been a useful aid in the management of sick neonates for the past few decades. However, it is associated with various complications. Reported studies strongly suggest a significant role of intravascular catheterization in the development of aortic thrombi. Increase in thrombosis of large vessels is believed to be related to mechanical injury in the catheterized vessels, which provide direct exposure of blood to tissue factor (TF), the primary cellular initiator of the extrinsic coagulation pathway. This study was conducted to determine the levels of plasma TF, tissue factor pathway inhibitor (TFPI) and D-dimer (DD) in infants with umbilical arterial catheter (UAC)-associated thrombosis. Quantification of TF was carried out using an in-house sandwich ELISA, whereas TFPI and DD levels were measured with commercial immunoassay kits. Infants with UAC inserted were found to have significantly higher levels of plasma TF (p < 0.001) than baseline levels. However, there were no significantly elevated levels of TFPI or DD. Infants with UAC-associated thrombosis demonstrated a greater increase of TF level (median: 414.5 pg/mL; range: -76.0, 6667.0) than infants without UAC-associated thrombosis (105.0 pg/mL; -976.0, 9480.0; p = 0.009) following UAC insertion. Our findings indicate that quantification and monitoring of TF levels could predict thrombus formation in infants with indwelling UAC. Following umbilical arterial catheterisation, infants with an approximately 3-fold rise in plasma TF levels were most at risk of developing abdominal aorta thrombosis as confirmed by real-time abdominal ultrasonography.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Fulltext Telomerase activation in neoplastic cell immortalization and tumour progression

Looi LM, Ng MH, Cheah PL

Malays J Pathol, 2007 Jun;29(1):33-5.
PMID: 19105326 MyJurnal

The unique ability of tumour cells to proliferate indefinitely is crucial to neoplastic progression as it allows these cells to express the aggressive properties of cancer without the censure of physiological ageing. This is in contrast to normal somatic cells which are subject to a "mitotic clock," a phenomenon that has been linked to telomeric shortening after each round of cell replication, so that eventually the loss of genetic material reaches a critical stage and the cells undergo senescence and cell death. A study was conducted to investigate the role of telomerase, an RNA-containing enzyme that restores the telomere length, in the neoplastic cell immortalization and progression process. Fresh human tissue samples taken from excision specimens received by the Department of Pathology, University of Malaya Medical Centre, were investigated for telomerase activity using a commercial Telomerase PCR-ELISA kit (Boehringer Mannheim). Specimens comprised 33 breast lesions (10 infiltrating breast adenocarcinoma, 13 fibroadenoma and 10 non-neoplastic breast tissue), 27 colonic lesions (17 colonic adenocarcinoma and 10 non-neoplastic colonic mucosa) and 42 cervical lesions (20 cervical carcinoma and 22 non-neoplastic cervical tissues). Telomerase activity was found in 6 (60%) of 10 breast carcinomas, 6 (46%) of 13 fibroadenomas, none of the 10 nonneoplastic breast samples, 3 (17.6%) of 17 colon carcinomas and none of the 10 non-neoplastic colonic mucosal samples, 12 (60%) of 20 cervical carcinoma and 3 (13.6%) of 22 non-neoplastic cervical samples. 5/10 (50%) Stage I, 4/7 (57%) Stage II, 2/2 (100%) Stage III and 1/1 (100%) Stage IV cervical carcinomas showed telomerase activity. These findings support a contributory role for telomerase in tumourigenesis with activation occurring from neoplastic transformation and increasing with tumour progression.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Fulltext Microalbuminuria measurements by two in-house ELISA methods

Ng LC, Teng LC, Ng ML, Sazali BS, Khalid BA

Malays J Pathol, 2000 Dec;22(2):73-8.
PMID: 16329538

Detection of microalbuminuria is important in the management of diabetic patients since it is predictive of development of proteinuria and nephropathy. Two sensitive and specific in-house ELISAs for microalbuminuria were established and validated. One of the ELISAs was based on antigen coating while the other employed antibody coating. Recovery and linearity experiments gave acceptable results of 100 +/- 10%, while precision results were <10% for intra-assay and <12% for inter-assay coefficients of variation (CVs). The standard curve ranged from 10-625 ug/l, equivalent to 0.2-12.5 mg/l for urine samples diluted 1:20 fold. When the antibody coated ELISA was compared to antigen coated ELISA, a correlation of r=0.996 was obtained. When compared to commercial kits, the in-house ELISAs gave good correlations of r=0.961 versus the Boehringer Mannheim Micral Test strips and r=0.940 versus Ames Microalb Turbidimetry. The normal microalbumin reference ranges determined for 12h, first morning and random urine samples were 0.7-5.3 mg, 0.1-10.2 mg/l and 0.8-26.1 mg/l respectively. The normal albumin excretion rate (AER) was 1.0-7.3 ug/min while untimed urine samples gave results of 0.1-0.9 and 0.2-1.6 mg/mmol after dividing by creatinine concentrations. The ELISAs were used to detect microalbuminuria in 338 random urine samples from diabetic patients. A high percentage 47.9% was found to be positive for microalbuminuria and 18.0% had macroalbuminuria >25 mg/mmol. Thus screening for microalbuminuria together with creatinine measurements using random urine samples can be used for management of diabetic patients.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*
Fulltext Increased soluble HLA-DRB1in B-cell acute lymphoblastic leukaemia

Hassan N, Dhaliwal JS, Mohd Ibrahim H, Osman R, HIdris SZ, Lee le J, et al.

Malays J Pathol, 2015 Aug;37(2):83-90.
PMID: 26277663 MyJurnal

Soluble HLA (sHLA) are potential tumour markers released in order to counter immune surveillance. sHLA-class II is less known especially in acute lymphoblastic leukaemia (ALL). This study aimed to investigate soluble, surface and allelic expression of HLA Class II (sHLA-DR) in B-cell ALL patients and compare with soluble expression in normal individuals. A sandwich enzyme-linked immunosorbent assay (ELISA) was developed to measure soluble HLA-DRB1 in plasma. Flow cytometric analysis was performed to determine median fluorescence intensity in HLA-DR surface expression. HLA-DNA typing by polymerase chain reaction, sequence specific oligonucleotides, PCRSSO was performed to determine HLA-DRB1 type in ALL samples. Results showed sHLA-DRB1 (mean±SEM) was significantly increased (p=0.001) in plasma of ALL patients (0.260 ±0.057 μg/mL; n=30) compared to healthy controls (0.051 ± 0.007µg/mL; n=31) of Malay ethnicity. However, these levels did not correlate with percentage or median fluorescence intensity of HLA-DR expressed on leukemia blasts (CD19+CD34 ± CD45(lo)HLA-DR+) or in the normal B cell population (CD19+CD34- CD45(hi)HLA-DR+) of patients. No significant difference was observed in gender (male/female) or age (paediatric/adult). Only a trend in reduced sHLA was observed in patients carrying HLA-DR04. These results have to be validated with a larger number of samples.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Detection of Legionella pneumophila antigens in patients' sera using monoclonal antibodies

Normaznah Y, Saniah K, Noor Rain A, Norazah A, Azizah MR, Sabiha P

Malays J Pathol, 1998 Dec;20(2):95-8.
PMID: 10879269

Three monoclonal antibodies (McAb) were produced against soluble antigens of Legionella pneumophila serogroup 1 which was cultured on BCYE agar. The McAbs were all of the IgM isotype. The McAbs were used in the McAb-based ELISA for detection of circulating L. pneumophila antigens in 186 sera collected from patients with symptoms and signs suggestive of atypical pneumonia. The normal reference optical (OD) density value of each of the McAbs was determined using 44 sera collected from healthy blood donors. The antigen positivity rates for the McAbs 1C7.2B, 2B2.10F and 2B2.11E were 11.3%, 7.7% and 22.2% respectively. Antigen positivity of the McAb 2B2.10F was significantly higher in the younger age group (p < 0.05). There is no significant association between the antigen positivity with age and sex for all the McAbs. There was no cross-reaction demonstrated between the McAbs with other bacterial antigens.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Detection of false positives by incorporation of a confirmatory blocking test into a commercial enzyme-linked immunosorbent assay specific for adenovirus antigen

Kirnpal-Kaur BS, Yap KL, Tan SC

Malays J Pathol, 1997 Dec;19(2):133-6.
PMID: 10879254

A blocking test was incorporated into the commercial IDEIA Adenovirus test (DAKO Diagnostics Ltd., Cambridgeshire, UK) to detect false positive results when faecal specimens were tested for adenovirus antigen. Immune rabbit serum raised against pooled adenovirus particles from human faecal specimens, together with the pre-immune serum, was used. Assessment of positive showed that false positives were produced under two different conditions: when results were based on visual determination instead of a cut-off value determined from photometric reading, and when absorbance values were not immediately read at the end of the test. Under the optimum condition for reading and assessment of test results (immediate reading and photometric determination), 11% of 65 adenovirus-positive samples were checked by the blocking ELISA as false positives. The rest of the specimens showed blocking of positive absorbance values by 70 to 98%. ELISA was found to be more sensitive than immune electron microscopy on samples with lower antigen concentration.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*
Detection of Chlamydia trachomatis in urine samples by polymerase chain reaction and enzyme immunoassay

Ngeow YF, Hema V, Zakaria M, Lee CH, Ramachandran S

Malays J Pathol, 1997 Dec;19(2):127-32.
PMID: 10879253

First-void urine samples collected from sexually transmitted diseases (STD) clinic patients were examined by a nested polymerase chain reaction (PCR) and a commercial enzyme immunoassay (IDEIA Chlamydia) for the diagnosis of Chlamydia trachomatis urethritis or cervicitis. The primers for the PCR amplified a target in the major outer membrane protein (MOMP) gene in C trachomatis while the IDEIA detected genus-specific chlamydial lipopolysaccharide. Discrepant results were resolved by retesting urine specimens with a second (plasmid-based) PCR and taking urethral or endocervical swab results into consideration. For 231 men (chlamydial prevalence 20.4%), the sensitivity, specificity, positive and negative predictive values were 59.6%, 99.5%, 96.6% and 90.6% for urine IDEIA, 68.1%, 99.5%, 97% and 92.4% for urethral swab IDEIA and 97.9%, 99.5%, 97.9% and 99.5% for urine PCR. The corresponding rates for 66 women (chlamydial prevalence 54.6%) were 19.4%, 100%, 100% and 50.8% for urine IDEIA, 86.1%, 96.7%, 96.9% and 85.3% for endocervical swab IDEIA and 91.7%, 93.3%, 94.3% and 90.3% for urine PCR. Hence, in a high prevalence population, the urine IDEIA was a suitable alternative to the male urethral swab IDEIA but significantly less sensitive than the endocervical swab IDEIA. The urine PCR was, however, much more sensitive than the urine IDEIA for both men and women and could replace the endocervical swab IDEIA for the diagnosis of chlamydial cervicitis.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
An evaluation of the staphylococcal co-agglutination test for the detection of group A rotavirus in human faeces

Yap KL, Ooi YE, Khor CM, Wong SH

Malays J Pathol, 1992 Dec;14(2):105-10.
PMID: 1338997

The group A rotavirus staphylococcal co-agglutination test was evaluated and its sensitivity and specificity compared with an in-house enzyme-linked immunosorbent assay (ELISA) and a commercial latex agglutination test (Rotalex). In addition, the storage stability of the staphylococcal reagents was ascertained. Examination of 136 clarified suspensions of diarrhoeal faeces by the staphylococcal co-agglutination test revealed a high proportion of false positives (26%) and uninterpretable results (34%) due to non-specific agglutination. Non-specific agglutination could be removed effectively by prior absorption of the clarified faecal specimens with unsensitized staphylococci. The staphylococcal co-agglutination test was less sensitive and specific than the in-house enzyme-linked immunosorbent assay but was comparable to the Rotalex slide latex agglutination test. The staphylococcal reagents have a shelf life of at least 29 weeks.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Surfactant protein A and stable microbubble formation in tracheal aspirates

Cheong KB, Cheong SK, Boo NY

Malays J Pathol, 1996 Dec;18(2):101-5.
PMID: 10879230

This study aimed to determine the role of surfactant protein A (SP-A) in the formation of stable microbubble in tracheal aspirates. Our results showed that as the concentration of anti SP-A antibodies added to tracheal aspirate specimens increased, the number of stable microbubble formed in the specimen decreased. The correlation between stable microbubble counts and the SP-A levels in the tracheal aspirates was good, r = 0.85, p < 0.05. This study suggests that SP-A plays an important role in stable microbubble formation. Measurement of small stable microbubbles is thus a useful bedside test for predicting the SP-A activity in the tracheal aspirates and in indirect measurement of lung maturity.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Application of the Serodia-HCV particle agglutination for the detection of antibodies to hepatitis C virus

Ooi BG, Sinniah M, Ismail S, Baharuddin R

Malays J Pathol, 1996 Dec;18(2):89-93.
PMID: 10879228

The Serodia-HCV Particle Agglutination (HCV-PA) for the detection of HCV antibodies was compared with the Enzyme Immunoassay Test (UBI HCV EIA) for possible in-house use. A total of 150 specimens were analysed using UBI HCV EIA and Serodia-HCV PA. Of these, 80 (53.3%) were both PA and EIA positive and 59 (39.3%) were negative by both techniques. Eleven sera (7.4%) were found to be EIA-positive but PA-negative. These 11 discordant sera were further tested by the LiaTek-HCV III Immunoassay (Organon Teknika). Ten were found to be line immunoassay negative and one was line immunoassay positive. Failure of the PA to detect the HCV positive serum meant that a small proportion of HCV antibody positives may be missed by the PA test. We conclude that (i) EIA should continue to be the first line screening test in our laboratory, (ii) PA with its 100% specificity could be a useful supplementary screen for all EIA-positive sera and finally (iii) line immunoassay could be used on sera to resolve discordant results in the EIA and PA assays.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Rapid dengue diagnosis and interpretation

Lam SK

Malays J Pathol, 1993 Jun;15(1):9-12.
PMID: 8277797
Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods
An in-house enzyme-linked immunoabsorbant assay for human growth hormone

Nazaimoon W, Ng ML, Bak K

Malays J Pathol, 1993 Jun;15(1):75-83.
PMID: 8277795

A simple, non-isotopic in-house enzyme-linked immunoabsorbant assay (ELISA) for human growth hormone (GH) was developed. The assay involved using in-house polyclonal anti-GH adsorbed onto 96-well microtitre plates, commercially prepared mouse monoclonal anti-GH, and goat anti-mouse IgG horseradish peroxidase detection system. Results of recovery and parallelism studies ranged from 95%-106% and 98%-101% respectively, of the expected values. The detection limit of the assay was 0.008 mIU/well or the equivalent to 0.4 mIU/L of undiluted serum. Intra- and interassay coefficients of variations were 4.8%-7.9% and 6.5%-8.7% respectively. Serum GH levels measured in this assay correlated well with those measured in established in-house radioimmunoassays (r = 0.985, p < 0.001) and immunoradiometric assay from NETRIA (r = 0.984, p < 0.001).

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*
An in-house ELISA method for measuring surfactant protein A

Cheong KB, Cheong SK, Boo NY, Jemilah M, Ton SH

Malays J Pathol, 1995 Dec;17(2):91-6.
PMID: 8935133

An in-house enzyme-linked immunoabsorbant assay (ELISA) for SP-A was successfully developed using in-house polyclonal anti SP-A and a commercial polyclonal anti-rabbit immunoglobulin horseradish peroxidase conjugate system. The standard curve, generated by using 50 ng of SP-A to coat the plate and 1:500 dilution of polyclonal anti SP-A as a primary antibody, was linear for concentrations of SP-A ranging from 4 micrograms/l to 4000 micrograms/l and reproducible. Results of recovery study of SP-A from a known sample of tracheal aspirate ranged from 94%-114%. Intra- and inter-assay coefficients of variations were 2.7% and 5.6% respectively for a known sample of tracheal aspirate. Interference study showed that tracheal aspirate did not interfere with the assay. The assay developed was intended to be used for SP-A measurement in tracheal aspirates obtained from neonates with and without respiratory distress syndrome.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*; Enzyme-Linked Immunosorbent Assay/standards
Fulltext Use of dengue blot in dengue diagnosis: the Malaysian experience

Fang R, Sinniah M, Kuen LS

Malays J Pathol, 1992 Dec;14(2):117-20.
PMID: 1304624

Dengue fever/Dengue haemorrhagic fever (DF/DHF) has been a public health problem in Malaysia with an endemic level of about 7 per 100,000 population per year. In 1990, Malaysia experienced its most severe outbreak of DF/DHF with a record total of 5,590 cases referred to the Division of Virology, Institute for Medical Research (IMR). Of these, 1,880 were confirmed serologically to be DF/DHF. The conventional serological procedure, the Haemagglutination Inhibition (HI) test, for the diagnosis of DF/DHF is cumbersome and causes delay in diagnosis. Another problem associated with the HI test has been that it has often been difficult to obtain a second convalescent serum sample for an accurate diagnosis. This has raised an urgent need to establish a "rapid" test for diagnosis of DF/DHF. As such the authors recently carried out an evaluation of a newly available commercial rapid test, namely, the Dengue Blot Assay (Diagnostic Biotechnology Singapore Pte Ltd). The test is intended for use in laboratory confirmation of dengue virus infection. The evaluation was to determine if the test could be utilised as a routine laboratory test and to establish its sensitivity and specificity. Over 400 samples were tested against the Dengue Blot Assay. Results were checked against an in-house Dengue IgM ELISA and HI assay. Preliminary results indicate that the sensitivity and specificity of the Dengue Blot is satisfactory. Our results also indicate that the Dengue Blot has a useful role to play in a routine laboratory especially since it provides rapid results on single serum samples thereby reducing the workload in a busy diagnostic laboratory.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay
Fulltext An in-house ELISA method for quantification of circulating tissue factor

Tay SP, Cheong SK

Malays J Pathol, 2002 Jun;24(1):45-51.
PMID: 16329555

Tissue Factor (TF) is a low molecular weight transmembrane glycoprotein that initiates the clotting protease cascade. It is considered to be the principal regulator of the extrinsic coagulation pathway, hemostasis and thrombosis, as well as inflammation and cellular immune response. An in-house two-step direct sandwich ELISA (enzyme-linked immunosorbent assay) for immunological quantification of plasma TF was successfully developed. The assay employed a monoclonal antibody against human TF (1:400 dilution; 1250 ng/ml) and peroxidase-conjugated anti-TF IgG (1:1000 dilution; 2000 ng/ml) as capture and detecting antibodies respectively, whilst tetramethylbenzidine/H2O2 were utilized as substrates. Titration curves of recombinant TF were linear within 10 to 4000 pg/ml, with a detection limit of 36.31 pg/ml. It demonstrated low intra- (2.50 - 9.23 CV%) and inter-assays (5.65 - 13.57 CV%) variability, as well as satisfactory analytical recovery (91.55 - 103.95%) and good parallelism. The assay developed was intended to be applied for measurement of plasma TF levels in patients with thrombotic disorders.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay/methods*
Fulltext Identification of Haptoglobin as a Potential Biomarker in Young Adults with Acute Myocardial Infarction by Proteomic Analysis

Mohamed Bakrim N, Mohd Shah ANS, Talib NA, Ab Rahman J, Abdullah A

Malays J Med Sci, 2020 Mar;27(2):64-76.
PMID: 32788843 MyJurnal DOI: 10.21315/mjms2020.27.2.8

Background: Acute myocardial infarction (AMI) molecular research in young adults is still limited. The aim of this study is to identify AMI proteomic biomarker(s) in young adults.
Methods: This study comprised of two phases namely discovery and verification. In the discovery phase, proteins in the pooled plasma samples from young male adults between 18 and 45 years (10 AMI patients and 10 controls) were separated using two-dimensional electrophoresis. The protein spots that were expressed differently in the AMI patients were identified via matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. The plasma concentrations of these proteins were quantified using enzyme-linked immunosorbent assay during the verification phase (40 AMI patients and 80 controls).
Results: Haptoglobin (Hp), apolipoprotein AI (Apo AI) and apolipoprotein AIV (Apo AIV) were up-regulated in the discovery phase. In the verification phase, the plasma concentration of Hp was significantly higher in AMI patients than the controls (P < 0.001). Logistic regression showed an association between Hp and AMI in young adults (odds ratio [OR] = 1.016, 95% CI: 1.002-1.030, P = 0.025) independent of other AMI risk factors. Hp was significantly correlated with high sensitivity C-reactive protein (hs-CRP) (r = 0.424, P < 0.001).
Conclusion: In young adults with AMI, plasma Hp concentrations were elevated and it is independently associated with AMI. A positive correlation with hs-CRP suggests Hp could be a potential biomarker of AMI in young adults.

Matched MeSH terms: Enzyme-Linked Immunosorbent Assay

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