Leptospira

Prof. Dr. Mochammad Hatta, Ph.D., Sp.MK (K)

Bagian Mikrobiologi
Fakultas Kedokteran,
Universitas Hasanuddin
Makassar

Mochammad Hatta@2013
 Leptospirosis Synonyms
Weils syndrome
Weils disease Swineherds disease
Hemorrhagic jaundice Cane cutters disease
Infectious jaundice Rice field workers disease
Yellows Hay makers disease
Seven day fever Fish handlers disease
Dairy farm fever
Field fever
Water fever Harvest fever
Mud fever Japanese autumnal
Swamp fever
Marsh fever Mouse fever
Canicola fever
Stuttgart disease Enzoonotic disease
Fort Bragg fever

Mochammad Hatta@2013
Cause and Manifestations of
Leptospirosis

Mild and flu-like

WEILS syndrome (jaundice, b
oliguria/anuria)
Meningitis
Haemorrhages (notably of lungs)

Mochammad Hatta@2013
Symptoms and Signs of Leptospirosis
Fever Oliguria/anuria
General malaise Jaundice
Headache Haemorrhages
Stiff neck Skin rash
Chills Conjunctival suffusion
Muscle pain Photophobia
Abdominal pain Cough
Joint pain Cardiac arrythmia
Anorexia Hypotension
Nausea Mental confusion
Vomitting Psychosis
Diarrhoea Dilirium

Mochammad Hatta@2013
Differential Diagnosis of Leptospirosis
Influenza
Dengue-virus infection (Dengue Hemorrhagic Fever)
Hanta-virus infection
Other viral disease causing haemorrhagic fevers
Rickettsiosis
Borreliosis
Brucellosis
Malaria
Pyelonephritis
Aseptic meningitis
Chemical poisoning
Food poisoning
Enteric-like diseases
Jaundice with fever (yellow fever, hepatitis)
Pyrexia of unknown origin (PUO)
Mochammad Hatta@2013
 Leptospirosis
Risk Groups
(Cattle) farmers
Rice field workers
Sugar cane cutters
Slaughterhouse workers
Sewer workers
Fisherman

Inhabitants of slum areas

Recreants
Persons involved in (traffic) accidents
Mochammad Hatta@2013
 Leptospirosis
Risk Factors
Animal factor
Investation with rodents
Infected domestic animals
Human factor
Poor hygiene
Low socioeconomic level
Abrased / cut skin
Environment
Warm and wet climate, pH of soil
Flooding
Mochammad Hatta@2013
 Causative Agent of
Leptospirosis
Pathogenic spirochaetes of the species
Leptospira interrogans (> 200 serovars)
Each serovar causes a similar clinical
picture
Icterohaemorrhagiae and copenhageni: Weils disease
lai: lung haemorrhagic form
grippotyphosa and hardjo: mild disease

Mochammad Hatta@2013
 Laboratory Diagnosis of
Leptospirosis
Culture, MAT and ELISA are standard
diagnostic tests for leptospirosis

Culture is slow
MAT and ELISA are complicated
Sensitivity depends on stage of illness

Microscopy of clinical specimens: non-

specific!
Mochammad Hatta@2013
 Sensitivity of Leptospirosis
Serology

Duration of the disease (days)

Percentage positive
1-3 4-6 7-9 10-12 13-15

MAT 33.3% 46.4% 58.3% 79.4% 96.7%

IgM ELISA 33.3% 67.9% 66.7% 74.4% 90.6%

Mochammad Hatta@2013
 LeptoTek lateral flow assay:
a practical new tool for diagnosing leptospirosis

Fast and reliable detection within 10 minutes

No special equipment or training required
Very user friendly
25 test devices per kit
Detects Leptospira-specific IgM antibodies in Control line
serum samples and whole blood Test line

Negative Positive

Mochammad Hatta@2013
 LEPTO Lateral Flow
Principle

Immunochromatographic strip assay

Test Control

Sample pad / Conjugate Detection strip Sink

blood cell pad
separation filter

Mochammad Hatta@2013
 LEPTO Lateral Flow
Method

Add 5l serum
Add 130l sample fluid
Control line
Wait 10 minutes Test line

Read result
Sample well

Mochammad Hatta@2013
 LEPTO Lateral Flow
Sensitivity and specificity

<10 DPO >10 DPO

LFA MAT ELISA LFA MAT ELISA
Sensitivity 65.9% 69.5% 56.7% 80.9% 93.6% 84.3%

Specificity 93.3% 98.6% 95.9% 96.2% 98.8% 98.8%

Mochammad Hatta@2013
 LEPTO Lateral Flow
Finger Prick Blood
Malankara Medical Mission Hospital
Kerala, India

Reciprocal IgM ELISA titre Lateral flow assay result

Negative Weak positive Positive

<20 131 5 1
20
40
80 4 5
160 1 1 9
320 5
640 14
1280 8
2560 12

94.3% agreement, kappa = 0.87 (95% CI, 0.74-1.00)

Mochammad Hatta@2013
 Laboratory Testing and
Clinical Diagnosis
Percentage positive in:
Clinical diagnosis IgM ELISA Lateral flow

Leptospirosis (n=96) 61% 63%

Leptospirosis or viral disease (n=19) 0% 0%

Viral disease (n=30) 0% 0%

Other (n=28) 0% 0%

Mochammad Hatta@2013
Rapid diagnostic test for leptospirosis

LeptoTek Lateral Flow Assay

Strong positive Weak positive Negative

- High Sensitivity & Specificity

- High Stability of Components
- No Requirement for Training and Special Equipment
Mochammad Hatta@2013
 LeptoTek Dri Dot:
The rapid screening assay for
leptospirosis
Fast and reliable detection within 30 sec
No special equipment or training required
Very user friendly
30 test devices per kit
Detects Leptospira-specific antibodies
Highly stable with shelf life over 2 years at +2C to +45

Negative Positive

Mochammad Hatta@2013
 LEPTO Dri Dot
Sensitivity and specificity

< 10 DPO > 10 DPO

Dri Dot MAT ELISA Dri Dot MAT ELISA

Sens 72.3% 65.5% 59.6% 88.2% 91.7% 89.3%

Spec 93.3% 98.9% 95.9% 89.8% 99.2% 99.3%

Mochammad Hatta@2013
 Test for Leptospirosis
Conclusions
Sensitivity and specificity similar to ELISA
Easy and rapid to use
Assay format suitable for field use
Available

In endemic areas results may vary due to

residual antibodies present of previous
exposure / cross-reacting antibodies
Mochammad Hatta@2013