OBJECTIVETo develop an up-converting phosphor technology-based lateral-flow (UPT-LF) assay for rapid quantitative detection of Burkholderia pseudomallei on site.

METHODSThe strip Bps-UPT-LF strip was prepared with up-converting phosphor (UCP) particles as the bio-label using double-antibody sandwich method. Detection performance, including sensitivity, quantitative accuracy, precision, and specificity, were first evaluated using bacterial suspensions of Burkholderia pseudomallei, the related species and the strains which had similar routes of transmission with serial standard concentrations diluted by phosphate buffer, then biological and chemical reagents and simulated samples with series concentrations were employed for sample tolerance evaluation, while the operation error during on site detection was also evaluated through adjusting liquid measure.

RESULTSThe whole detection was accomplished within 20 minutes, and the sensitivity was 10(4) CFU/ml with linear quantitative range from 10(4) CFU/ml to 10(7) CFU/ml, which covered four orders of magnitude. Bps-UPT-LF strip demonstrated high specificity with the absence of any false-positive result even at 10(7) and 10(8) CFU/ml of non-specific bacterial contamination. Not only Bps-UPT-LF strip could tolerate to high concentration of the extreme acid and basic matter (pH 1-12), saline matter (≤ 2 mol/L mixture of NaCl and KCl), viscous materials (≤ 50 g/L of PEG 20000 and ≤ 20% of glycerol) and bio-macromolecule (≥ 400 g/L of bovine serum albumin or ≥ 80 g/L of casein), but also it can directly detect animal, environmental and powder specimen, such as ≥ 400 g/L of milk powder, flour powder, fruit juice, fresh and decomposed viscera, and ≤ 200 g/L of putty powder, sucrose, gourmet powder, and soil. Operation errors of liquid measure had few effects on sensitivity and specificity, including -50%-200% of sample, -22%-44% of sample-treating buffer and -30%-30% of loading mixture.

CONCLUSIONThe good detection performance and tolerance performance bring the bright future for Bps-UPT-LF strip to detect Burkholderia pseudomallei on site rapidly and quantitatively for nature foci surveillance and anti-bioterrorism.

BACKGROUND: In the search for more powerful tools for diagnoses of endemic diseases in resource-limited settings, we have been analyzing technologies with potential applicability. Increasingly, the process focuses on readily accessible bodily fluids combined with increasingly powerful multiplex capabilities to unambiguously diagnose a condition without resorting to reliance on a sophisticated reference laboratory. Although these technological advances may well have important implications for the sensitive and specific detection of disease, to date their clinical utility has not been demonstrated, especially in resource-limited settings. Furthermore, many emerging technological developments are in fields of physics or engineering, which are not readily available to or intelligible to clinicians or clinical laboratory scientists. CONTENT: This review provides a look at technology trends that could have applicability to high-sensitivity multiplexed immunoassays in resource-limited settings. Various technologies are explained and assessed according to potential for reaching relevant limits of cost, sensitivity, and multiplex capability. Frequently, such work is reported in technical journals not normally read by clinical scientists, and the authors make enthusiastic claims for the potential of their technology while ignoring potential pitfalls. Thus it is important to draw attention to technical hurdles that authors may not be publicizing. SUMMARY: Immunochromatographic assays, optical methods including those involving waveguides, electrochemical methods, magnetorestrictive methods, and field-effect transistor methods based on nanotubes, nanowires, and nanoribbons reveal possibilities as next-generation technologies.
Endemic Diseases ; Health Resorts ; Immunoassay ; Immunochromatography ; Nanotubes ; Nanotubes, Carbon ; Nanowires

BACKGROUND: Broth cultures are increasingly used to detect acid-fast bacilli (AFB). Rapid, simple and accurate methods for differentiation of Mycobacterium tuberculosis complex (MTBC) and nontuberculosis mycobacteria from broth cultures are needed. Immunochromatographic assays (ICTs) for identification of MTBC have been developed. METHODS: The abilities of the BD MGIT TBc Identification Test (Becton Dickinson, USA) and the SD Bioline TB Ag MPT64 (Standard Diagnostics, Korea) to detect MTBC were evaluated in 44 AFB-positive broth cultures. The results of 2 ICTs were compared to those of real-time PCR. RESULTS: The BD MGIT TBc Identification Test and the SD Bioline TB Ag MPT64 showed concordant results with real-time PCR by 100% and 97.7%, respectively. The sensitivity of the BD MGIT TBc Identification and the SD Bioline TB Ag MPT64 was 100% for both, and the specificities of those were 100% and 95.2%, respectively. CONCLUSIONS: Both ICTs are rapid methods for identification of MTBC from broth cultures, and the results of ICTs are in accord with those of real-time PCR.
Immunochromatography ; Mycobacterium ; Mycobacterium tuberculosis ; Real-Time Polymerase Chain Reaction

BACKGROUND: For the diagnosis of tuberculosis (TB), a variety of tests based on the patients' immune response has been introduced. We evaluated the clinical usefulness of combined anti-tuberculosis antibody (anti-TB Ab) test and Interferon-gamma release assay (IGRA), evaluating humoral and cellular immune response to Mycobacterium tuberculosis, respectively. METHODS: Among patients tested for IGRA, 78 patients diagnosed as TB and treated with anti-TB drug and 80 non-TB patients were included in this study. We used QuantiFERON-TB GOLD (QFT, Cellestis limited, Australia) for IGRA and an immunochromatographic assay, Easy Test TB (ASAN PHARM, Korea), for anti-TB Ab test. RESULTS: The sensitivity, specificity, and positive and negative predictive values of Easy Test TB were 23.1%, 98.8%, 94.7% and 56.8%, respectively. QFT had a significantly higher sensitivity than Easy Test TB (67.9% vs. 23.1%; P<0.05). The agreement between the two assays was poor (69.6%, k=0.190). Of the 18 cases with positive Easy Test TB, six (33%) showed negative QFT results. The combination of Easy Test TB and QFT had a significantly higher sensitivity than single QFT (75.6%, vs. 67.9%; P=0.031). CONCLUSIONS: The combination of Easy Test TB and QFT could be used to aid in a rapid diagnosis and early treatment of TB.
Humans ; Immunity, Cellular ; Immunochromatography ; Interferon-gamma ; Interferon-gamma Release Tests ; Mycobacterium tuberculosis ; Sensitivity and Specificity ; Tuberculosis

OBJECTIVES: The purpose of this study was to suggest a proper method for the detection of heaptitis B surface antibody(anti-HBs) in a screening program for hepatitis B vaccination. METHODS: Sensivitity, specificity and predictive values were compared between Immunochromatographic assay (ICA) and passive hemagglutination(PHA) in 978 subjects(565 males, 413 females, 19-78 years ranging in age, mean 46.5 years old). EIA was used as a standard method for the detection of HBsAb. RESULTS: Sensitivity in the detection of anti-HBs of PHA and ICA was 88.7% and 94.9%, specificity was 94.3% and 96.6%, negative predictive value was 96.5% and 98.0%, and positive predictive value was 82.3% and 91.3%, respectively. False negative rate(11.3%) of PHA was higher than that(5.1%) of ICA. The higher the titer of anti-HBs in EIA was, the lower the false negative rate was. There was no false negative result in the cases with 101mIU/ml or more in EIA. CONCLUSION: We suggest that ICA should be the choice of screening method in the detection of anti-HBs in Hepatitis B vaccination program.
Female ; Hepatitis B* ; Hepatitis* ; Humans ; Immunochromatography ; Korea* ; Male ; Mass Screening ; Sensitivity and Specificity ; Vaccination*

The followings are the results for external quality assessment (EQA) in immunoserology for 2004: 1. Evaluation of EQA was done in 2 trials in May and November, about 99% of laboratories participating average 8.4 items. EQA for anti-HBc test was newly started in 2004. 2. Commercial control, MASR Immunology Control from Medical Analysis Systems (Camarillo, CA, USA) was used to assure the quality of quantitative results of C-reactive protein (CRP), rheumatoid factor (RF) and anti-streptolysin O (ASO) tests in 2004. All the specimens for Immunoserology in EQA were delivered refrigerated for the first time, being received within 48 hours after sending. 3. EQA for detection of HBsAg mutants was tried for the first time, using the recombinant HBsAg mutant (Gly/Arg 145) kindly provided by Abbott Laboratories, USA. 4. The laboratories using immunochromatography assay (ICA) were increased, however, many laboratories using ICA reported falsely negative for the positive specimens. The sensitivity of ICA test kits as well as various factors influencing the ICA results should be evaluated. 5. Standardization of methods including calibrators for quantitative results should be required for the harmonization of results.
Allergy and Immunology ; C-Reactive Protein ; Hepatitis B Surface Antigens ; Immunochromatography ; Korea* ; Nephelometry and Turbidimetry ; Rheumatoid Factor

Mycobacterium tuberculosis complex (MTBC) is discriminated from non-tuberculous mycobacteria (NTM) via an immunochromatographic assay (ICA) which is based on the reactions of monoclonal antibodies against MPT64, one of the predominant proteins excreted by MTBC. Recently, the authors of the present study discovered SD TB-negative Mycobacterium tuberculosis strains. In addition, sequence analysis of the mpt64 genes in these strains was performed and showed a deletion of 63 bp from nucleotides 196 to 258. In cases of MPT64-negative mycobacterium, the authors recommend performing TB PCR for correct diagnosis.
Antibodies, Monoclonal ; Immunochromatography ; Mycobacterium ; Mycobacterium tuberculosis ; Nucleotides ; Polymerase Chain Reaction ; Proteins ; Sequence Analysis

BACKGROUND: To detect active tuberculosis, clinicians usually rely on several methods those are so limited. As the prevalence rate of tuberculosis is high in Korea, culture is not so very effective in clinical use. The polymerase chain reaction (PCR) featuring rapidness and high sensitivity offers low specificity and it requires high test cost, complicated skills, expensive equipment. This study attempted to determine if the immunochromatographic assay, intended to measure antibodies using 38 kDa antigens, is valuable as a new method to diagnose active tuberculosis, by comparing it with existing acid-fast stain (AFB stain) and PCR. METHODS: The sera were collected from 31 BCG-vaccinated healthy persons and 55 patients subjected to AFB stain and PCR who visited Pohang Hospital of Dongguk University or Kyungpook National University Hospital, and then kept at -20degrees C until experiment. Fifty-five patients subjected to AFB stain and PCR were composed of 24 active tuberculosis patients and 31 non-TB patients. The evaluation of active tuberculosis was based on clinical criteria. RESULTS: The detection rate of antibody by the immunochromatographic method accounted for 83% in the active TB group, and each 6% in both the non-TB group and the healthy control group. The sensitivities of AFB stain, PCR and immunochromatographic method accounted for 67%, 88% and 83%, respectively, the specificities for 94%, 86% and 94%, respectively, the positive predictive values for 89%, 84% and 91%, respectively, and the negative predictive values for 78%, 89% and 88%, respectively. CONCLUSIONS: This suggests that the immunochromatographic method can be used for the rapid diagnostic method of active tuberculosis in an area with high prevalence value of tuberculosis like Korea. In addition, the immunochromatographic method showed the sensitivity approximate to that of PCR, the same specificity as AFB stain, and a high positive and negative predictive values. So it was expected not only to be greatly helpful for the diagnosis of active tuberculosis but also to be more useful in clinical practices because of short examination time, no special equipment and skills required, and inexpensive examination.
Antibodies ; Diagnosis* ; Gyeongsangbuk-do ; Humans ; Immunochromatography* ; Korea ; Polymerase Chain Reaction ; Prevalence ; Sensitivity and Specificity ; Tuberculosis*

BACKGROUND: This study was conducted to evaluate the usefulness of the BACTEC MGIT (Mycobacterium Growth Indicator Tube) 960 system for mycobacteria culture and immunochromatographic assay to identify Mycobacterium tuberculosis (MTB) in positive MGIT culture. METHODS: Mycobacteria-culture-positive cases were retrospectively analyzed from December 2010 to July 2011. The detection rates and the recovery times of the mycobacteria between the Ogawa media and the MGIT were compared. An immunochromatographic assay (ICA) (SD BIO-LINE) was also performed in the positive MGIT culture for identification, and the results were compared with those of the Ogawa media in the Korea National Tuberculosis Association. RESULTS: Among the 261 patients (M:F, 168:93; mean age, 61.6+/-17.16 yrs), 450 specimens (sputa, 365; bronchial washing, 61; and pleural effusion, 24) were found positive with mycobacteria. Mycobacteria were grown both on the MGIT and Ogawa media in 310 cases (68.9%); only on the MGIT in 115 cases (22.6%); and only on the Ogawa media in 25 cases (5.5%) (p<0.05).The recovery time was 28.2+/-8.9 days in the Ogawa media and 11.1+/-5.8 days in the MGIT (p<0.05). Among the 127 cases from the positive MGIT culture, all 92 cases that were confirmed as MTB cases bythe Korea National Tuberculosis Association were identified as MTB by ICA, with 100% sensitivity. CONCLUSION: MGIT increases the detection rate and shortens the recovery time of mycobacteria in clinical respiratory specimens, and the TB Ag MPT64 kit using ICA is useful in identifying MTB in a positive MGIT culture.
Humans ; Immunochromatography ; Korea ; Mycobacterium ; Mycobacterium tuberculosis ; Pleural Effusion ; Retrospective Studies ; Tuberculosis

BACKGROUND: Many diagnostic tests have developed to diagnose tuberculosis and other mycobacterial diseases but the diagnosis of tuberculosis relies largely on radiological findings and acid-fast staining of sputum and/or culture. Recently, new serologic diagnostic methods, which are safe and easy to use have been introduced into Korea. In this study, the usefulness of serologic diagnosis for tuberculosis and the disease pattern induced variation of the test were evaluated. METHODS: Serological assay was performed upon 108 patients with two test kits, the ICT tuberculosis and the BioSign(TM) TB, which are based upon a rapid immunochromatographic assay technique, capable of being interpreted within 15 minutes. The case groups consisted of 61 patients with active pulmonary tuberculosis(36 patients), extrapulmonary tuberculosis(3 patients), or both (22 patients). Control groups consisted of 47 patients with inactive old pulmonary tuberculosis (17 patients), nontuberculous pulmonary disease(16 patients) and nonpulmonary cardiac disease(14 patients). RESULTS: The diagnostic sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the ICT tuberculosis were 64.3%, 91.5%, 90.0% and 68.3% respectively. The diagnostic sensitivity, specificity, PPV and NPV of the BioSign(TM) TB were 76.5%, 95.3%, 94.1% and 78.8% respectively. Differences in sensitivity were not significant between patients with previous history of tuberculosis or patients without prior history of tuberculosis. The ICT tuberculosis test showed higher sensitivity in pulmonary tuberculosis patients (76.5%) than extrapulmonary tuberculosis patients (33.3%). There was no difference in sensitivity between patients with or without cavitary lesion by chest X-ray. CONCLUSION: Considering high specificity and PPV, serologic diagnosis using a rapid immunochromatographic assay device is another helpful diagnostic method in the diagnosis of tuberculosis, when combined with previous diagnostic methods such as chest X-ray, microbiologic study but it has limitation in terms of confirming the diagnosis for tuberculosis as the only diagnostic method because of relatively low sensitivity and NPV.
Diagnosis* ; Diagnostic Tests, Routine ; Humans ; Immunochromatography* ; Korea ; Sputum ; Thorax ; Tuberculosis* ; Tuberculosis, Pulmonary