Diagnostic test for leptospirosis

Instead of clinical symptoms to ensure that someone suffering leptospirosis

can safety some test to confirm the diagnosis,the test can be divided into microscopic
demonstration,culture

and

serology.In

culture

test

there

are

isolation

of

leptospires,identification of leptospiral isolates and susceptibility testing.Serology test

can divided into enzyme-linked immunosorbent assay (ELISA) and microscopic
agglutination test(MAT)[1]
Microscopic demonstration is to view leptospires in cell,leptospires may be
visualized in clinical material by dark field microscopy or by immunofluorence or
light microscopy after appropriate staining.Microscopy of blood is of value only
during the first few days of the acute illness while leptospiremia occurs.Staining
methods have been applied to increase the sensitivity of direct microscopic
examination.These include immunofluorescence staning of bovine urine,water,and
soil immunoperoxidase staining of blood and urine [1].Leptospires were first visualized
by silver staining,and the warthin-starry stain is widely used for histologic
examination.
Culture test for leptospirosis can be mention such as isolation of
leptospires,identification of leptospiral isolates and susceptibility testing.Isolation of
leptospires is the early test for diagnose leptospirosis,blood cultures should be taken
as soon as possible after the patients presentation,to process the test patient must
drops of blood are inoculated into 10 ml of semisolid medium containing 5flurouracil at the patient bedside[1].Inoculation of media with dilutions of blood
samples may increase recovery,leptospires survive in conventional blood culture
media for number of days.Isolation of leptospires from clinical samples gives
definitive diagnosis and also aids in identifying the prevalent serovar.Urine also can
be also cultured from the beginning of the second week of symptomatic
illness,survival of leptospires in voided human urine is limited,so urine should be
process immediately by centrifugation,followed by resuspending the sediment in

phosphate buffered saline and inoculating into ng into semisolid medium containing
5-fluoruracil.Culture are incubated at 28o to 30o and examined weekly by dark field
microscopy for up to 13 weeks before being discarded[2].Identification of leptospiral
is task for specialized reference laboratories.For complete identification,a
combination of procedures is used to determine 1) if the isolate is a pathogen or a
saprophyte; 2) the species of Leptospira to which the isolate belongs; and 3) the
serogroup and serovar of the isolate.Isolated leptospires are identified by serological
methods or by recently developed molecular techniques [2],which can perform the
microscopic agglutination test to identify isolates with relative rapidity.Susceptibility
testing is the test to know leptospires susceptibility with.Leptospires are susceptible
to

beta-lactams,macrolides,tetracyclines,fluoroquinolones

and

streptomycin[1]

.Problem in the determination of susceptibility include the long incubation time

required,the use of media containing serum and the difficulty in quantifying growth
accurately.
Serological testing is the laboratory procedure most frequently used to
confirm the clinical diagnosis, to determine herd prevalence, and to conduct
epidemiological studies. Leptospiral antibodies appear within a few days of onset of
illness and persist for weeks or months and, in some cases, years. The MAT using live
antigens is the most widely used serological testThe MAT using live antigens is the
most widely used serological test,after incubation,the serum antigen mixtures are
examine microscopically for agglutination and the titers determined [3].The MAT is
read by darkfield microscopy,the end point is the highest dilution of serum at which
50%agglutination occurs,because of the difficulty in detecting when 50%of the
leptospires are agglutinated,the end point is the presence of approximately 50% free.
[1]

immunoglobulin M enzyme-linked immunosorbent assay (IgM ELISA) is

promising because it can be performed in a greater number of laboratories throughout

the tropics, and is inexpensive compared with MAT. IgM antibodies become
detectable during the first week of illness allowing the diagnosis to be confirmed and
treatment initiated while it is likely to be most effective [1].Antibody levels are
generally low or absent during very early infection.

1. Health M, Army I. Laboratory Diagnosis of Leptospirosis. 2005;51(3):195200.

2. Manual OT. Leptospirosis. Version Adopt by World Assem Deleg OIE un May
2014.2014;(May):1-15.
3. Baburaj P, Nandkumar VS, Khanna L. Polymerase chain reaction in the

diagnosis

of

leptospiral

2006;54(April):339-340.

infection.

Assoc

Physicians

India.