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Chapter 61 : The and

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Abstract:

The order consists of the family and the family . The family contains the genus and the genus . Members of the family are short rods or coccobacilli, and members of the family are small pleomorphic cocci. The family consists of two genera, one of which, , includes human pathogens belonging to the α2-group proteobacteria. Currently, the principal techniques used for the serodiagnosis of rickettsial diseases are probe-based immunoassays. Immunoprobe-based tests include the indirect fluorescence assay (IFA) and its micromodification, the micro-IF (MIF) test. The MIF test is still considered as the gold standard. Other immunoprobe tests include immunoperoxidase assays (IPAs); enzyme-linked immunosorbent assays (ELISAs), in which the antigens are adsorbed either onto the well of a microtiter plate or onto nitrocellulose in a dot blot or slot blot configuration; and immunoblot assays (IBAs). IFAs and IPAs require the whole bacterium as the antigen. The remaining immunoprobe assays use either highly purified rickettsiae or an extract. The development of real-time PCR-based diagnosis and rapid PCR-based diagnostic methods has been the focus of much attention in many research laboratories. Recently, a protein-based method for the detection of the small-cell variant protein A (ScvA) plus other protein markers comprising a fingerprint via matrix-assisted laser desorption-time of flight (MALDI-TOF) mass spectrometry has been developed.

Citation: Hechemy K, Rikihisa Y, Macaluso K, Burgess A, Anderson B, Thompson H. 2006. The and , p 526-539. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch61

Key Concept Ranking

RNA Polymerase beta Subunit
0.5131579
Rocky Mountain Spotted Fever
0.44334602
Restriction Fragment Length Polymorphism
0.4124688
0.5131579
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Figures

Image of FIGURE 1
FIGURE 1

Taxonomy of human pathogens of the order with representative species from each genus and bacteria from genera that were formerly in the order and were kept in this chapter for historical purposes.

Citation: Hechemy K, Rikihisa Y, Macaluso K, Burgess A, Anderson B, Thompson H. 2006. The and , p 526-539. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch61
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Image of FIGURE 2
FIGURE 2

Correlation between IFA titers and ODs for ELISA with EK-rP44 as the antigen. Each circle represents one sample. The rho value calculated by Spearman’s rank correlation was 0.740 ( <0.001; 181). (Reprinted with permission from reference .)

Citation: Hechemy K, Rikihisa Y, Macaluso K, Burgess A, Anderson B, Thompson H. 2006. The and , p 526-539. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch61
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Image of FIGURE 3
FIGURE 3

(A) Dot blot immunoassay of HGE patient sera with different IFA testing titers by use of 0.5 μg of affinity-purified recombinant P44 (rP44) antigen. The sera were diluted 1:1,000. (B) Layout of experiment. (Reprinted with permission from reference .)

Citation: Hechemy K, Rikihisa Y, Macaluso K, Burgess A, Anderson B, Thompson H. 2006. The and , p 526-539. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch61
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Image of FIGURE 4
FIGURE 4

Western immunoblot analysis of anti-HGE (presently, anti-HGA) sera using purified HGE (presently, HGA) agent isolate 13 and rP44 antigen. The sera used for this study included a horse anti-HGE-agent serum; five convalescent-phase serum samples, from patients 2, 3, 11, 13, and 16; five serum samples collected at different times of illness over a 2-year period from patient 21, who was suspected of having persistent infection or reinfection; and negative control serum (IFA titer, <1:20). Samples subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis consisted of 10 μg of purified whole-cell preparations of HGE agent strain 13, affinity-purified rP44, HL-60 cells, and BL21(DE30)/pLysS. These proteins were transferred to a nitrocellulose sheet and incubated with a 1:1,000 dilution of antisera. The number at the bottom of each panel represents the IFA test titer of the serum sample. The numbers on the left indicate molecular masses in kilodaltons based on broad-range prestained standards (Bio-Rad) (adapted and reprinted with permission from reference ).

Citation: Hechemy K, Rikihisa Y, Macaluso K, Burgess A, Anderson B, Thompson H. 2006. The and , p 526-539. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch61
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Tables

Generic image for table
TABLE 1

Representative types of pathogenic rickettsiae in humans

Citation: Hechemy K, Rikihisa Y, Macaluso K, Burgess A, Anderson B, Thompson H. 2006. The and , p 526-539. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch61
Generic image for table
TABLE 2

Examples of PCR primer sets that allow for identification and differentiation of rickettsial species via RFLP analysis

Citation: Hechemy K, Rikihisa Y, Macaluso K, Burgess A, Anderson B, Thompson H. 2006. The and , p 526-539. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch61
Generic image for table
TABLE 3

Primers used for PCR

Citation: Hechemy K, Rikihisa Y, Macaluso K, Burgess A, Anderson B, Thompson H. 2006. The and , p 526-539. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch61

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