Further evaluation of one-point microcapsule agglutination test for diagnosis of leptospirosis

Cui, Jie; Xiao, Guozhi; Chen, T. Z.; Zhu, Genfeng; Sato, T.; Seki, Masaaki; Kobayashi, Seiichi; Arimitsu, Yoshiko

doi:10.1017/s0950268800067625

Cited by 5 publications

(5 citation statements)

References 3 publications

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“…A microcapsule agglutination test using a synthetic polymer in place of red blood cells (39, 514) has been evaluated extensively in Japan and China (40,139). In an international multicenter evaluation, the microcapsule agglutination test was more sensitive than either the MAT or an IgM-ELISA in early-acute-phase samples (38), but failed to detect infections caused by some serovars (38,513).…”

Section: Downloaded Frommentioning

confidence: 99%

Leptospirosis

2001

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Leptospirosis is a worldwide zoonotic infection with a much greater incidence in tropical regions and has now been identified as one of the emerging infectious diseases. The epidemiology of leptospirosis has been modified by changes in animal husbandry, climate, and human behavior. Resurgent interest in leptospirosis has resulted from large outbreaks that have received significant publicity. The development of simpler, rapid assays for diagnosis has been based largely on the recognition that early initiation of antibiotic therapy is important in acute disease but also on the need for assays which can be used more widely. In this review, the complex taxonomy of leptospires, previously based on serology and recently modified by a genotypic classification, is discussed, and the clinical and epidemiological value of molecular diagnosis and typing is also evaluated

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Section: Downloaded Frommentioning

confidence: 99%

Leptospirosis

2001

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“…For example, for patients 1, 2, and 10, the highest A f values and MAT titers were for serogroup icterohaemorrhagiae, whereas for patients 3,5,7,8,9,15,16,17, and 18 the highest values were for serogroup hardjo.…”

Section: Determination Of Leptospiral Agglutination By Fcmmentioning

confidence: 95%

“…Among them are the complement fixation test (33), several enzyme-linked immunosorbent assay formats (1,29), the macroscopic slide agglutination test (14), the microcapsule agglutination test (9), the indirect hemagglutination assay (20), the dipstick assay (27), and other methods (3,15,22,30). Each assay has its own advantages, drawbacks, and limitations (4,18).…”

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confidence: 99%

New Approach for Serological Testing for Leptospirosis by Using Detection of Leptospira Agglutination by Flow Cytometry Light Scatter Analysis

Yitzhaki¹,

Barnea²,

Keysary³

et al. 2004

J Clin Microbiol

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Leptospirosis is considered an important reemerging infectious disease worldwide. The standard and most widespread method for the diagnosis of leptospirosis is the microscopic agglutination test (MAT). This test is laborious and time-consuming, and the interpretation of the results is subjective. In the present work we describe an application of flow cytometry (FCM) as a tool for the serological diagnosis of leptospirosis. The analysis is based on the sensitivity of FCM to the size and shape of the bacteria analyzed by measurement of light scatter parameters: forward scatter (FSC) and side scatter (SSC). The addition of positive serum to an infecting leptospiral serovar results in a shift of the light scatter parameter to a different location with higher FSC and SSC values, indicating the formation of leptospiral aggregates. By using immunofluorescent staining, we have shown that the large particles formed are the agglutinated leptospires. Quantification of the agglutination process has been achieved by calculating an agglutination factor (A f ), based on changes in the light scatter parameters measured by FCM. A f enables us to determine the specificity of the serological reaction of the patient serum with each leptospiral serovar. In this work, 27 serum samples from 18 leptospirosis patients were tested by both the MAT and the FCM techniques, in which each serum sample was tested against 13 serovars. Twenty-six human serum samples derived from patients with a variety of other defined illnesses were used as negative controls and enabled us to define the A f threshold value as <9.3 for negative patients, while any value higher than that would be a positive result for leptospirosis. Compared to MAT, the FCM technique was found to be more specific and sensitive, especially in identifying the serogroup in the acute phase of the disease. The whole process was found to be rapid and took less than 1.5 h. Moreover, FCM analysis is objective and can be automated for the handling of large numbers of samples.

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“…Complement fixation test (Wolff, 1954), Enzyme linked immunosorbent assay (Vanasco et al, 2001), the macroscopic agglutination test (Galton et al, 1958), the microcapsule agglutination test (Cui et al, 1991), the indirect hemagglutination assay (Levett and Whittington, 1998) the dipstick assay (Smits et al, 1999) are some of the methods and other methods (Torten et al, 1966). The Polymerase chain reaction based diagnostic tests for leptospirosis are speed, sensitive and an indication of current infection status (Zuerner and Bolin, 1997;Zuerner et al, 1995) each assay has its own advantages, drawbacks and limitations Levett, 2001).…”

Section: Introductionmentioning

confidence: 99%

Characterization, Comparison Study of Outer Membrane Protein OMPL1 of Pathogenic Leptospira Species for Disease Diagnosis

Muthiah¹,

Kumar²,

Ramadass³

2015

Research J. of Microbiology

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Leptospirosis is an acute bacterial infection which affects humans and a wide range of animals this bacterial disease is caused by pathogenic members of the genus Leptospira. It is now known that this is only one of more than 200 pathogenic serovars distributed between 7 species and 23 serogroups worldwide. Commercial leptospiral vaccines rely on bacterins and inactivated whole cells. A major component of immunity resulting from whole cell vaccines is a humoral immune response resulted against the serovar specific carbohydrate antigen of leptospiral lipopolysaccharide. Cross protection against many of the 250 serovars of pathogenic Leptospira species is lacking though the immunity generated is readily demonstrable. Hence there is an urgent need for development of an improved leptospiral vaccine. Variations in the carbohydrate side chains of LPS are responsible for the antigenic diversity observed among Leptospiral serovars. The present study was undertaken to develop a recombinant fusion protein from Leptospira as antigen by the following processes. Identification and purification of gene encoding the leptospiral outer membrane OMPL1 by PCR. Cloning and expression of the OMPL1 gene in a suitable vector system. Purification and characterization of the expressed recombinant leptospiral outer membrane protein OMPL1. Standardization of recombinant antigen based diagnostic study.

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Further evaluation of one-point microcapsule agglutination test for diagnosis of leptospirosis

Cited by 5 publications

References 3 publications

Leptospirosis

Leptospirosis

New Approach for Serological Testing for Leptospirosis by Using Detection of Leptospira Agglutination by Flow Cytometry Light Scatter Analysis

Characterization, Comparison Study of Outer Membrane Protein OMPL1 of Pathogenic Leptospira Species for Disease Diagnosis

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