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      Rapid Detection of Yersinia Pestis Antigen from Decomposed Rodent Viscera Using An Up-converting Phosphor Technology-Based Lateral-Flow Assay

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          Rapid quantitative detection of Yersinia pestis by lateral-flow immunoassay and up-converting phosphor technology-based biosensor

          Up-converting phosphor technology (UPT)-based lateral-flow immunoassay has been developed for quantitative detection of Yersinia pestis rapidly and specifically. In this assay, 400 nm up-converting phosphor particles were used as the reporter. A sandwich immumoassay was employed by using a polyclonal antibody against F1 antigen of Y. pestis immobilized on the nitrocellulose membrane and the same antibody conjugated to the UPT particles. The signal detection of the strips was performed by the UPT-based biosensor that could provide a 980 nm IR laser to excite the phosphor particles, then collect the visible luminescence emitted by the UPT particles and finally convert it to the voltage as a signal. V T and V C stand for the multiplied voltage units for the test and the control line, respectively, and the ratio V T/V C is directly proportional to the number of Y. pestis in a sample. We observed a good linearity between the ratio and log CFU/ml of Y. pestis above the detection limit, which was approximately 104 CFU/ml. The precision of the intra- and inter-assay was below 15% (coefficient of variation, CV). Cross-reactivity with related Gram-negative enteric bacteria was not found. The UPT-LF immunoassay system presented here takes less than 30 min to perform from the sample treatment to the data analysis. The current paper includes only preliminary data concerning the biomedical aspects of the assay, but is more concentrated on the technical details of establishing a rapid manual assay using a state-of-the-art label chemistry.
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            Early diagnosis of bubonic plague using F1 antigen capture ELISA assay and rapid immunogold dipstick.

            Plague is still prevalent in more than 20 countries. Two F1 antigen diagnostic assays (an immunocapture ELISA and an immunogold chromatography dipstick) were evaluated using bubo aspirates, serum and urine specimens from patients suspected with plague. The specificity of the two F1 assays was found 100%. Using bacteriology as a gold reference diagnostic assay, 52 patients were Yersinia pestis culture positive and 141 negative. The sensitivity of the F1 ELISA test was 100% in bubo, 52% in serum and 58% in urine specimens. In culture negative patients, the F1 antigen could be found in 10% bubo aspirates, 5% serum and 7% urine specimens of culture negative patients for whom a seroconversion for anti-F1 antibodies was also observed. The sensitivity of the dipstick assay was 98% on bubo aspirates specimens. Compared to the ELISA test, the agreement rate was 97.5% and the correlation coefficient tau = 0.90 (p < 10(-3)). In conclusion, the diagnosis of bubonic plague has to be performed on bubo fluid rather than on serum or urine specimens. Both the F1 ELISA and the dipstick assays are valuable tools for an early diagnosis and for the surveillance of plague.
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              Prospective evaluation of commercial antibody-based rapid tests in combination with a loop-mediated isothermal amplification PCR assay for detection of Orientia tsutsugamushi during the acute phase of scrub typhus infection.

              Samples from 160 prospectively recruited febrile patients with typhus-like illness in an area of Thailand (Chiang Rai, northern Thailand) where scrub typhus is endemic were used to evaluate the diagnostic capabilities of four rapid immunochromatographic tests (ICTs) for the detection of Orientia tsutsugamushi IgM and total antibodies during acute scrub typhus infection. Of the 160 cases, 54 (34%) had been confirmed to have scrub typhus using the reference scrub typhus infection criteria (STIC), i.e., positive cell culture isolation, an admission IgM antibody titer of ≥1:12,800, a 4-fold rising IgM antibody titer, and/or positivity for ≥2 out of 3 PCR gene targets). The ICTs gave the following sensitivities and specificities: the Panbio IgM ICT, 46% (95% confidence interval [CI], 33 to 60) and 95% (95% CI, 89 to 98), respectively; the Standard Diagnostics IgM ICT, 68% (95% CI, 60 to 75) and 73% (95% CI, 68 to 78), respectively; the AccessBio IgM ICT, 56% (95% CI, 48 to 63) and 90% (95% CI, 87 to 94), respectively; and the AccessBio total antibody ABt ICT, 61% (95% CI, 53 to 68) and 68% (95% CI, 63 to 73), respectively. An isothermal loop amplification (LAMP) PCR assay for scrub typhus demonstrated a sensitivity of 52% (95% CI, 38 to 66) and a specificity of 94% (95% CI, 88 to 98). This study has revealed the diagnostic limitations of antibody-based assays in an acute care setting. However, the combination of ICTs with LAMP usually increased sensitivity with a minimal reduction in specificity. The best combination, the Panbio IgM ICT and LAMP, resulted in a sensitivity of 67% (95% CI, 53 to 79) and a specificity of 91% (95% CI, 83 to 95). The combination of antibody-based assays with DNA- or antigen-based tests shows promise for improved diagnostic sensitivity.
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                Author and article information

                Contributors
                Journal
                Infectious Diseases and Translational Medicine
                Infect. Dis. Transl. Med.
                Infect. Dis. Transl. Med.
                International Biological and Medical Journals Publishing House Co., Limited (Room E16, 3/f, Yongda Commercial Building, No.97, Bonham Stand (Sheung Wan), HongKong )
                2411-2917
                30 December 2015
                : 1
                : 2
                : 58-60
                Affiliations
                From Laboratory of Analytical Microbiology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China; Beijing Key Laboratory of Point-of-Care Testing for Bioemergency and Clinic (No. BZ0329), Beijing 100071, China
                Department of Etiology, Taishan Medical University, Taian 271000, China; Clinical Diagnostic Center, 302nd Hospital of the People’s Liberation Army, Beijing 100039, China
                Qinghai Institute for Endemic Disease Prevention and Control, Xining 811602, China
                Clinical Diagnostic Center, 302nd Hospital of the People’s Liberation Army, Beijing 100039, China
                Qinghai Institute for Endemic Disease Prevention and Control, Xining 811602, China
                From Laboratory of Analytical Microbiology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China; Beijing Key Laboratory of Point-of-Care Testing for Bioemergency and Clinic (No. BZ0329), Beijing 100071, China
                Qinghai Institute for Endemic Disease Prevention and Control, Xining 811602, China
                Qinghai Institute for Endemic Disease Prevention and Control, Xining 811602, China
                Department of Etiology, Taishan Medical University, Taian 271000, China
                Qinghai Institute for Endemic Disease Prevention and Control, Xining 811602, China
                From Laboratory of Analytical Microbiology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing 100071, China; Beijing Key Laboratory of Point-of-Care Testing for Bioemergency and Clinic (No. BZ0329), Beijing 100071, China
                Author notes
                Correspondence to: Ailian Yu: Department of Etiology, Taishan Medical University, Taian 271000, China; Tel: 86-538-6222493; Fax: 86-538-6222493; Email: alyu@ 123456tsmc.edu.cn .
                Baizhong Cui: Qinghai Institute for Endemic Disease Prevention and Control, Zongzhai Town, Huangzhong City, Xining 811602, China; Tel: 86-971-6510197; Fax: 86-971-6510197; Email: cbzxn1@ 123456163.com .
                Lei Zhou: Laboratory of Analytical Microbiology, State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, No. 20, Dongdajie, Fengtai District, Beijing 100071, China; Tel: 86-10-66948562; Fax: 86-10-63815689; Email: ammszhoulei@ 123456aliyun.com .

                : These authors contributed equally to this work.

                Article
                10.11979/idtm.201502003
                1098bf94-66ec-4e99-944c-8d2abfcd6b88

                This work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 Unported License. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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                Figures: 0, Tables: 3, References: 13, Pages: 3
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                Categories
                Letter: Detection Techniques

                Medicine,Infectious disease & Microbiology
                Medicine, Infectious disease & Microbiology

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