Modification of the TUBEX typhoid test to detect antibodies directly from haemolytic serum and whole blood

2008 ◽  
Vol 57 (11) ◽  
pp. 1349-1353 ◽  
Author(s):  
Frankie C. H. Tam ◽  
Danny T. M. Leung ◽  
C. H. Ma ◽  
Pak-Leong Lim
Keyword(s):  
Typhoid Fever ◽  
Magnetic Separation ◽  
Whole Blood ◽  
Magnetic Particles ◽  
Original Method ◽  
Serum Antibodies ◽  
Assay Sensitivity ◽  
Simple Modification ◽  
Colour Changes ◽  
Larger Sample

The TUBEX test for typhoid fever detects serum antibodies in a simple and rapid assay system based on the inhibition of binding between two types of reagent particles — magnetic particles coated with an antigen (Salmonella O9 LPS) and coloured indicator particles coated with an anti-O9 mAb. A magnet is used to separate the colour indicator particles bound to the magnetic particles from the unbound indicator particles. Specific colour changes following magnetic separation are indicative of antibodies in the patient's serum; however, because results are interpreted based on changes in the colour red, haemolytic or icteric specimens cannot be used. This study describes a simple modification of the protocol to accommodate such specimens, including whole blood. This involves the addition of a quick and simple washing step after mixing the specimen with the antigen-bound magnetic particles. This modification has the advantage of allowing larger sample volumes to be used, thus enhancing the assay sensitivity, and also enables cases considered to be borderline positive by the original method to be re-assessed.

scholarly journals Direct magnetic separation of immune cells from whole blood using bacterial magnetic particles displaying protein G

2009 ◽  
Vol 25 (1) ◽  
pp. 219-226 ◽  
Author(s):  
Masayuki Takahashi ◽  
Tomoko Yoshino ◽  
Haruko Takeyama ◽  
Tadashi Matsunaga
Keyword(s):  
Magnetic Separation ◽  
Immune Cells ◽  
Whole Blood ◽  
Magnetic Particles ◽  
Protein G ◽  
Bacterial Magnetic Particles

Improved Nitromethane-Hyamine Method for the Chemical Determination of Nitrofurantoin in Whole Blood

1970 ◽  
Vol 16 (10) ◽  
pp. 820-823 ◽  
Author(s):  
G. L Mattok ◽  
I.J McGilveray ◽  
Claude Charette
Keyword(s):  
Whole Blood ◽  
Original Method ◽  
Chemical Determination

Abstract The nitromethane-Hyamine method for assay of nitrofurantoin in whole blood has been improved: smaller volumes of blood (0.8 ml) are required, the sensitivity is greater (0.2 pg/mI), and the solution to be read is more stable than in the original method. The analysis detects 70 to 72% of 1 to 4 Ag of nitrofurantoin added per milliliter of whole blood. Blood concen-trations after the usual therapeutic peroral dose (100 mg) of nitrofurantoin are reported.

Whole-Blood Amino Acids in Experimentally Induced Typhoid Fever in Man

1968 ◽  
Vol 278 (6) ◽  
pp. 293-298 ◽  
Author(s):  
Ralph D. Feigin ◽  
Albert S. Klainer ◽  
William R. Beisel ◽  
Richard B. Hornick
Keyword(s):  
Amino Acids ◽  
Typhoid Fever ◽  
Whole Blood ◽  
Experimentally Induced

CpG oligonucleotides increase HBV-specific cytokine responses in whole blood and enhance cytokine release assay sensitivity

2017 ◽  
Vol 248 ◽  
pp. 195-201 ◽  
Author(s):  
Werner Dammermann ◽  
Julia Dornbrack ◽  
Katharina Bröker ◽  
Frank Bentzien ◽  
Stefan Lüth
Keyword(s):  
Whole Blood ◽  
Cytokine Release ◽  
Assay Sensitivity ◽  
Cytokine Responses ◽  
Cpg Oligonucleotides ◽  
Release Assay

Improvement and assessment of enzyme-linked immunosorbent assay to detect low FK506 concentrations in plasma or whole blood within 6 hours

1993 ◽  
Vol 39 (6) ◽  
pp. 1045-1049 ◽  
Author(s):  
P E Wallemacq ◽  
I Firdaous ◽  
A Hassoun
Keyword(s):  
Detection Limit ◽  
Whole Blood ◽  
Clinical Investigation ◽  
Plasma Concentrations ◽  
Original Method ◽  
Enzyme Linked Immunosorbent Assay ◽  
Pharmacokinetic Studies ◽  
Blood Concentrations ◽  
Blood Temperature ◽  
Biological Variables

Abstract FK506, a promising new immunosuppressant, is currently under clinical investigation. Because dose-dependent toxicity is possible, blood concentrations of FK506 should be monitored. We improved the original ELISA of FK506 by shortening the incubation time. With some modification of materials, results are obtained within 6 h instead of 2 days, with similar or even better precision. Internal and external quality-control programs showed that our results correlated satisfactorily both with values determined by the original method and the theoretical expected values. Either plasma (detection limit 0.1 microgram/L) or whole-blood (detection limit 1 microgram/L) samples can be used. The sensitivity of the method makes it particularly useful for accurate pharmacokinetic studies or measurement of low blood concentrations. Twenty-four drugs and nine biological variables showed no significant interference on the assay. Study of the concentration- and temperature-dependent distribution of FK506 shows that the drug is largely bound to erythrocytes (ratio of blood to plasma concentrations is 10-40); as the erythrocytes become saturated, more of the drug is found in the plasma. Plasma concentrations may vary according to the blood temperature. We conclude that whole blood should be used for FK506 monitoring, as it is for monitoring cyclosporine.

Quantification and molecular profiling of circulating tumor cells (CTCs) in urothelial cancer (UC) before and during systemic treatment: Implications across the clinical stages.

2014 ◽  
Vol 32 (4_suppl) ◽  
pp. 319-319 ◽  
Author(s):  
Andrea Necchi ◽  
Emanuela Fina ◽  
Patrizia Giannatempo ◽  
Maurizio Colecchia ◽  
Chiara Iacona ◽  
...  
Keyword(s):  
Whole Blood ◽  
Urothelial Cancer ◽  
Clinical Relapse ◽  
Clinical Stages ◽  
Tgfβ Receptor ◽  
Stepwise Reduction ◽  
Muscle Invasive ◽  
Larger Sample ◽  
Combined Technique

319 Background: CTCs from patients (pts) at different clinical stages were analyzed by a never explored experimental approach based on a combination of two techniques. Provision of this information may contribute to optimize tailored therapies. Methods: 3 cohorts were analyzed: pts with muscle-invasive bladder cancer receiving neoadjuvant (NA) sorafenib + chemotherapy (CT) (NCT01222676), and metastatic (M1) pts receiving 1st-line MVAC, and 2nd-line (M2) anti-TGFβ receptor ALK1 PF-03446962 (NCT01620970). 5 ml of whole blood were filtered by ScreenCell Cyto devices and CTC status was assessed with centralized scoring by referral pathologists. Additional 5 ml of whole blood were processed by immunomagnetic beads (AdnaTestSelect kit) and the expression level of a panel of markers (including EPCAM and MUC1) was studied using RT-multiplex PCR. The objective was the association with clinical response/relapse. Results: From 07/2012 to 08/2013, 48 pts (17 NA, 17 M1, and 14 M2) were enrolled. Rates of baseline CTC+ were: 91, 77, 75% (ScreenCell), and 36, 41, 69% with AdnaTest, respectively. NA setting: from baseline to post-CT, all pts had a stepwise reduction of CTC count/5 ml blood by ScreenCell (median baseline of 15 [0-40] to 0 [0-3]). Increase in circulating EPCAM±MUC1 levels by CTC was seen in accordance with the 3 disease progressions (PD). M1 setting: Pts who relapsed had a median of 25 CTC/5 ml (IQR: 12-47) at the end of CT, while all the others had levels <10. EPCAM profile was concordant in all cases (median 1.10 vs 0.42 ng/µl). Despite classified as partial/complete responders by RECIST criteria, 5/7 cases with >6 month follow-up displayed an increase in EPCAM levels and CTC number which anticipated a relapse. M2 setting: increase in CTC signals from baseline to +2 months was observed in 8/9 evaluable cases by AdnaTest and 4/5 evaluable cases by ScreenCell, in accordance with PD. Conclusions: This combined technique showed a promising ability to anticipate the detection of clinical relapse, although it still needs a larger sample size. Refining molecular characterization will help designing informed clinical trials.

Washing-free Chemiluminescence Immunoassay for Rapid Detection of cTnI in Whole Blood Samples

2021 ◽  
Author(s):  
Huan Zhao ◽  
Enben Su ◽  
Li Huang ◽  
Yunfeng Zai ◽  
Yuan Liu ◽  
...  
Keyword(s):  
Whole Blood ◽  
Rapid Detection ◽  
Magnetic Particles ◽  
Troponin I ◽  
Sample Type ◽  
Serum Samples ◽  
Chemiluminescence Immunoassay ◽  
Blood Samples ◽  
Significant Difference ◽  
Whole Blood Samples

Abstract Background: Chemiluminescence immunoassay (CLIA) has always been a great challenge in detecting whole blood samples without centrifugation because of the interference of red blood cells and low sensitivity. Results: In this scheme, the antigens and erythrocytes in the blood were captured by the antibodies immobilized on the magnetic particles, recognized by another biotin-conjugated cTnI antibody and detected by streptavidin/acridine aster-conjugated PCMS. After magnetic separation, the supernatant was transferred and measured. No significant difference was noted between the cTnI concentrations of the serum samples, plasma samples and whole blood. The prepared PCMS provided more functional areas to conjugate streptavidin and acridinium ester, so the immunoassay has highly sensitive, the limits of blank at 0.012 ng/mL, and functional sensitivity at 0.019 ng/mL with a CV of 20%, and 0.058 ng/mL with a CV of 10%. Total precision of any sample type ranged from 2.62~5.67%. The assay was linear over the studied range of 0.01–50.00 ng/mL, and no hook effect was found when cTnI concentrations reached 1900 ng/mL. No significant interference was noted with the potential endogenous interfering substances. Compared with the commercial kit (Abbott assay kit), the correlation coefficient was 0.9859.Conclusions: A washing-free chemiluminescence immunoassay (CLIA) was established for the rapid detection of cardiac troponin I (cTnI) in human whole blood, using erythrocyte capture antibodies-conjugated magnetic nanoparticles for eliminating the influence of erythrocytes and polychloromethylstyrene microspheres (PCMS) for signal amplification, which showed great potential in clinical application.

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