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New Approach for Serological Testing for Leptospirosis by Using Detection

New Approach for Serological Testing for Leptospirosis by Using Detection

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Published by: TAMIL on Aug 10, 2009
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 New Approach for Serological Testing for Leptospirosis by Using Detection of 
Agglutination by Flow Cytometry Light Scatter Analysis
What are leptospires?
Leptospires are bacteria which can be either pathogenic (i.e. having thepotential tocause disease in animals and humans) or saprophytic (i.e. free living andgenerallyconsidered not to cause disease).Pathogenic leptospires are maintained in nature in the renal tubules of certainanimals.Saprophytic leptospires are found in many types of wet or humidenvironments rangingfrom surface waters and moist soil to tap water. Saprophytic halophilic (salt-loving)leptospires are found in seawater.
What is the medical significance of saprophytic leptospires?
 The saprophytes are supposed not to cause disea. Saprophytic leptospiresmay be found in cultures whensterility was not maintained during the preparation of culture media, whennon-sterileingredients were used for the preparation of culture media, or when clinicalsampleswere not collected aseptically.
Can pathogenic leptospires be distinguished from one another?
 The basic systematic unit is the serovar, defined on the basis of antigenicsimilarities and differences as revealed by the so-called cross-agglutinationabsorptiontest .Each serovar has a characteristic antigenic make-up.
Are there many serovars?
Serovars having antigenic similarities are formed into serogroups, and over200pathogenic serovars divided into 25 serogroups have been described.
What is the practical significance of the serovar concept?
. A certain serovar may develop a commensal orcomparatively mild pathogenic relationship with a certain animal hostspecies. Forinstance, cattle are often associated with serovar hardjo, dogs with canicolaand ratswith icterohaemorrhagiae and copenhageni.
What is the nature of the antibody response?
Humans react to an infection by leptospires by producing specific anti-
antibodies.Seroconversion may occur as early as 5–7 days after the onset of disease butsometimes only after 10 days or longer (see Clinical samples, p.9 andDetection of antibodies, p.10).IgM class antibodies usually appear somewhat earlier than IgG classantibodies, andgenerally remain detectable for months or even years but at low titreDetection of IgG antibodies is more variable. They may sometimes not bedetected atall, or be detectable for only relatively short periods of time, but maysometimes persistfor several years. The antibodies are directed against (
 Annex 8
common antigens (so-called genus-specific antigens) that are shared by allleptospires, both pathogenic and saprophytic;
serovar-specific and serogroup-specific antigens.Patients with leptospirosis may produce antibodies that react with severalserovars. This phenomenon, called cross-reaction, is often observed in the initial phaseof thedisease.After the acute disease, cross-reactive antibodies gradually disappear as theimmuneresponse "matures", usually in the course of weeks or months, whileserogroup- andserovar-specific antibodies often persist for years. Thus the genus-specific antibodies usually remain detectable for months, theserovarspecificantibodies for years.Weak cross-reactions may also occur to other groups of microorganisms andwill varywith the serological method used.Occasionally patients produce specific antibodies that react with broadlyreactiveantigens only, and few or no serovar-specific antibodies are detected. Someotherpatients may produce only serovar-specific antibodies.
Is the antibody response protective?
It is generally believed that serovar-specific antibodies are protective andthat a patientis immune to reinfection with the same serovar as long as the concentration(titre) of specific antibodies is high enough. Antibodies provoked by an infection with aparticular
serovar do not necessarily protect against infection with other serovars
What clinical samples should be collected for examination?
 This will depend on the phase of the infection. Leptospires usually circulate inthe bloodof the patient for about 10 days after the onset of the disease. They alsoappear inother body fluids, such as urine and cerebrospinal fluid, a few days after theonset of disease and penetrate internal organs during this time. Detectable titres of antibodiesappear in the blood about 5–10 days after the onset of disease, butsometimes later,especially if antibiotic treatment is instituted. The samples that are useful and most commonly collected are therefore:
Blood with heparin (to prevent clotting) for culture (
 Annex 12
) in the first10 days.Blood culture more than 10 days after disease onset is not worth while asleptospires have mostly disappeared from the blood and antibodies will havebecome detectable in the serum allowing serodiagnosis. Samples for cultureshould be stored and transported at ambient temperatures, since lowtemperaturesare detrimental to pathogenic leptospires.
Clotted blood or serum for serology (
 Annexes 10
). These shouldpreferablybe collected twice at an interval of several days based on the date of onset of disease and the probable time of seroconversion. The testing of paired sera isnecessary to detect a rise in titres between the two samples orseroconversion,and thus to confirm the diagnosis of leptospirosis (see Significance of serology,p.10). A negative serological result in the early phase of the disease does notexclude leptospirosis.
What laboratory method is most frequently used to diagnoseleptospirosis?
Current methods for the direct detection (
 Annexes 6, 12, 13
) of leptospires areeither slow or of limited reliability so that serology is often the mostappropriatediagnostic method. Moreover, in practice, patients often seek medical care orareadmitted in hospitals when they have already been ill for a sufficiently longtime to haveproduced detectable antibodies.
Is positive serology proof of a current infection?

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