18B - SEROLOGICAL AND MOLECULAR DETECTION OF SPIROCHETE DISEASES - LYME DISEASE AND LEPTOSPIROSIS

Borrelia burgdorferi

Loosely coiled spirochete

Outer surface proteins

Borrelia burgdorferi

Allow attachment to mammalian cells

OspA to OspF

Borrelia burgdorferi

OSPs from _ to _

Barbour-Stoenner-Kelly medium

Culture medium used to culture Borrelia burgdorferi

White footed mouse (Peromyscus leucopus)

Borrelia burgdorferi

Reservoir

Ixodes spp.

Borrelia burgdorferi

Vector

Penicillin, tetracycline (doxycycline), macrolide

Borrelia burgdorferi

Treatment (3)

Lyme disease

Borrelia burgdorferi

Disease

Localized Rash Stage

Early Dissemination Stage

Late Dissemination Stage

Three Stages of Lyme Disease

Localized Rash Stage

Three Stages of Lyme Disease

Hallmark of infection with Borrelia burgdorferi is the appearance of erythema migrans

erythema migrans

Three Stages of Lyme Disease

Hallmark of infection with Borrelia burgdorferi is the appearance of _ _

erythema migrans

Three Stages of Lyme Disease

Localized Rash Stage

Starts as a small red papule at the site of inoculation

erythema migrans

Three Stages of Lyme Disease

Localized Rash Stage

Expands to a ring-like erythema with partial clearing at the central area

“Bull’s eye rash”

Three Stages of Lyme Disease

Characteristic lesion in the Localized Rash Stage

Localized Rash Stage

Three Stages of Lyme Disease

Most serology results are negative at this stage

Early Dissemination Stage

Three Stages of Lyme Disease

Hematogenous dissemination of the organism

Early Dissemination Stage

Three Stages of Lyme Disease

Affects the skin, nervous system, heart, or joints

Early Dissemination Stage

Three Stages of Lyme Disease

Pain may also be present due to the dissemination of the organism

Facial palsy

Three Stages of Lyme Disease

Most prevalent neurological sign associated with Lyme disease

Late Dissemination Stage

Three Stages of Lyme Disease

Develops when Lyme disease is left untreated

Late Dissemination Stage

Three Stages of Lyme Disease

Presents with arthritis, peripheral neuropathy, and encephalomyelitis

two-tiered

clinical evidence

Laboratory Diagnosis for Lyme Disease

Uses a _-_ approach for diagnosis

Testing should be done with the presence of _ _

Standard Testing Algorithm

Laboratory Diagnosis for Lyme Disease

Patients are screened with EIA (ELISA) or IFA

Standard Testing Algorithm

Laboratory Diagnosis for Lyme Disease

A positive screening test is followed by Western Blot

Standard Testing Algorithm

Laboratory Diagnosis for Lyme Disease

Highly specific but low sensitivity in diagnosis of early Lyme disease

Modified Two-Tiered Algorithm

Laboratory Diagnosis for Lyme Disease

Uses ELISA with purified Borrelia peptide antigens

Modified Two-Tiered Algorithm

Laboratory Diagnosis for Lyme Disease

A positive screening test is followed by another EIA/ELISA with a different method

Western Blot

Laboratory Diagnosis for Lyme Disease

Confirmatory test for samples that initially test positive or equivocal by EIA or IFA

Western Blot

Laboratory Diagnosis for Lyme Disease

Consists of electrophoresis of Borrelia antigens in an acrylamide gel followed by transfer of the resulting pattern to nitrocellulose paper

IgM

Western Blot

2 out of 3 bands to confirm a positive result

23, 39, 41 kDa

Western Blot

3 bands for IgM

23 kDa

Western Blot

outer surface protein C band

41 kDa

Western Blot

flagellin band

IgG

Western Blot

5 out of 10 bands to confirm a positive result

18, 23, 28, 30, 39, 41, 45, 58, 66, 93 kDa

Western Blot

10 bands for IgG

Leptospirosis

Caused by Leptospira spp.

Leptospira spp.

Thin, flexible, and tightly coiled spirochetes with hook-like ends

chain

cocci

Spirals of Leptospira resemble a _ of _

mucous membranes, abrasions, and ingestion

Leptospira spp. is transmitted through humans by exposure via (3)

Fletcher, Ellinghausen-McCullough-Johnson-Harris (EMJH), or Stuart medium

Culture medium used for Leptospira spp. (3)

penicillin and doxycycline

Treatment for Leptospira spp. (2)

Leptospira spp.

S/S will present abruptly and non-specific

Weil’s Disease

severe form of Leptospirosis which involves a severe systemic disease (kidneys and liver)

Darkfield

low sensitivity

Laboratory Diagnosis for Leptospirosis

_ microscopy can be used in the first week of illness but is not recommended due to _ _

Serology

Laboratory Diagnosis for Leptospirosis

most common method to diagnose leptospirosis

Microscopic Agglutination Test (MAT)

Laboratory Diagnosis for Leptospirosis

gold standard for the diagnosis of Leptospirosis

Microscopic Agglutination Test (MAT)

Laboratory Diagnosis for Leptospirosis

All positive screening tests must be confirmed by