Treponema pallidum particle agglutination assay

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Fig. 1: Microwells showing positive and negative TPHA test TPHA test on microplate wells.jpg
Fig. 1: Microwells showing positive and negative TPHA test

The Treponema pallidum particle agglutination assay (also called TPPA test) is an indirect agglutination assay used for detection and titration of antibodies against the causative agent of syphilis, Treponema pallidum subspecies pallidum. It also detects other treponematoses.[ citation needed ]

In the test, gelatin particles are sensitized with T. pallidum antigen. Patient serum is mixed with the reagent containing the sensitized gelatin particles. The particles aggregate to form clumps when the patient serum is positive for syphilis. In other words, the patient's serum contains antibodies to T. pallidum. A negative test shows no clumping of gelatin particles. This is a type of specific treponemal test for syphilis. [ citation needed ]

A similar specific treponemal test for syphilis is the Treponema pallidum hemagglutination assay or TPHA. TPHA is an indirect hemagglutination assay used for the detection and titration of antibodies against the causative agent of syphilis, Treponema pallidum subspecies pallidum.[ citation needed ]

In the test, red blood cells (erythrocytes) are sensitized with antigens from T. pallidum pallidum. The cells then aggregate on the surface of a test dish if exposed to the serum of a patient with syphilis. It is used as a confirmatory test for syphilis infection. A negative test result shows a tight button or spot of red blood cells on the surface of the test dish. Often a plastic test plate containing many small "wells" is used as the test dish so that many patients may be tested at the same time but their results can be kept separate from each other. Please note the image (Fig 1) of the wells on the test plate and the positive and negative test results look different. [1]

For primary syphilis, TPPA has a sensitivity of 85% to 100%, and a specificity of 98% to 100%. [2] In secondary and late-latent syphilis, TPPA has a sensitivity of 98% to 100%. [2]

Antibodies against other treponemal organisms, such as the T. pallidum subspecies endemicum, pertenue, or carateum, can cause false positive results. Not all these disease are venereal; it has been recommended that a careful explanation of this fact be included with test results. [3] A variety of methods exist to adsorb these antibodies from the test sample before hemagglutination.[ citation needed ]

Related Research Articles

<span class="mw-page-title-main">Syphilis</span> Sexually transmitted infection

Syphilis is a sexually transmitted infection caused by the bacterium Treponema pallidum subspecies pallidum. The signs and symptoms of syphilis vary depending in which of the four stages it presents. The primary stage classically presents with a single chancre though there may be multiple sores. In secondary syphilis, a diffuse rash occurs, which frequently involves the palms of the hands and soles of the feet. There may also be sores in the mouth or vagina. In latent syphilis, which can last for years, there are few or no symptoms. In tertiary syphilis, there are gummas, neurological problems, or heart symptoms. Syphilis has been known as "the great imitator" as it may cause symptoms similar to many other diseases.

<i>Treponema pallidum</i> Species of bacterium

Treponema pallidum, formerly known as Spirochaeta pallida, is a spirochaete bacterium with various subspecies that cause the diseases syphilis, bejel, and yaws. It is transmitted only among humans. It is a helically coiled microorganism usually 6–15 μm long and 0.1–0.2 μm wide. T. pallidum's lack of either a tricarboxylic acid cycle or oxidative phosphorylation results in minimal metabolic activity. The treponemes have a cytoplasmic and an outer membrane. Using light microscopy, treponemes are visible only by using dark-field illumination. T. pallidum consists of three subspecies, T. p. pallidum, T. p. endemicum, and T. p. pertenue, each of which has a distinct associated disease.

<span class="mw-page-title-main">Yaws</span> Medical condition

Yaws is a tropical infection of the skin, bones, and joints caused by the spirochete bacterium Treponema pallidum pertenue. The disease begins with a round, hard swelling of the skin, 2 to 5 cm in diameter. The center may break open and form an ulcer. This initial skin lesion typically heals after 3–6 months. After weeks to years, joints and bones may become painful, fatigue may develop, and new skin lesions may appear. The skin of the palms of the hands and the soles of the feet may become thick and break open. The bones may become misshapen. After 5 years or more, large areas of skin may die, leaving scars.

<span class="mw-page-title-main">Rapid plasma reagin</span> Test for syphilis

The rapid plasma reagin test is a type of rapid diagnostic test that looks for non-specific antibodies in the blood of the patient that may indicate an infection by syphilis or related non-venereal treponematoses. It is one of several nontreponemal tests for syphilis. The term reagin means that this test does not look for antibodies against the bacterium itself, Treponema pallidum, but rather for antibodies against substances released by cells when they are damaged by T. pallidum. Traditionally, syphilis serologic testing has been performed using a nontreponemal test (NTT) such as the RPR or VDRL test, with positive results then confirmed using a specific treponemal test (TT) such as TPPA or FTA-ABS. This algorithm is currently endorsed by the U.S. Centers for Disease Control and Prevention (CDC). In addition to screening for syphilis, a titer can be used to track the progress of the disease over time and its response to therapy. The traditional algorithm using an NTT followed by a TT remains the standard in many parts of the world.

<span class="mw-page-title-main">Venereal Disease Research Laboratory test</span>

The Venereal Disease Research Laboratory test (VDRL) is a blood test for syphilis and related non-venereal treponematoses that was developed by the eponymous US laboratory. The VDRL test is used to screen for syphilis, whereas other, more specific tests are used to diagnose the disease.

<span class="mw-page-title-main">Albert Ludwig Sigesmund Neisser</span> German physician

Albert Ludwig Sigesmund Neisser was a German physician who discovered the causative agent (pathogen) of gonorrhea, a strain of bacteria that was named in his honour.

Serology is the scientific study of serum and other body fluids. In practice, the term usually refers to the diagnostic identification of antibodies in the serum. Such antibodies are typically formed in response to an infection, against other foreign proteins, or to one's own proteins. In either case, the procedure is simple.

A Coombs test, also known as antiglobulin test (AGT), is either of two blood tests used in immunohematology. They are the direct and indirect Coombs tests. The direct Coombs test detects antibodies that are stuck to the surface of the red blood cells. Since these antibodies sometimes destroy red blood cells, a person can be anemic and this test can help clarify the condition. The indirect Coombs detects antibodies that are floating freely in the blood. These antibodies could act against certain red blood cells and the test can be done to diagnose reactions to a blood transfusion.

<span class="mw-page-title-main">Nonvenereal endemic syphilis</span> Medical condition

Bejel, or endemic syphilis, is a chronic skin and tissue disease caused by infection by the endemicum subspecies of the spirochete Treponema pallidum. Bejel is one of the "endemic treponematoses", a group that also includes yaws and pinta. Typically, endemic trepanematoses begin with localized lesions on the skin or mucous membranes. Pinta is limited to affecting the skin, whereas bejel and yaws are considered to be invasive because they can also cause disease in bone and other internal tissues.

<span class="mw-page-title-main">Agglutination (biology)</span> Type of antibody response

Agglutination is the clumping of particles. The word agglutination comes from the Latin agglutinare.

Hemagglutination, or haemagglutination, is a specific form of agglutination that involves red blood cells (RBCs). It has two common uses in the laboratory: blood typing and the quantification of virus dilutions in a haemagglutination assay.

<span class="mw-page-title-main">Neurosyphilis</span> Infection of the central nervous system in a patient with syphilis

Neurosyphilis is the infection of the central nervous system in a patient with syphilis. In the era of modern antibiotics, the majority of neurosyphilis cases have been reported in HIV-infected patients. Meningitis is the most common neurological presentation in early syphilis. Tertiary syphilis symptoms are exclusively neurosyphilis, though neurosyphilis may occur at any stage of infection.

The fluorescent treponemal antibody absorption (FTA-ABS) test is a diagnostic test for syphilis. Using antibodies specific for the Treponema pallidum species, such tests would be assumed to be more specific than non-treponemal testing such as VDRL but have been shown repeatedly to be sensitive but not specific for the diagnosis of neurosyphilis in cerebrospinal fluid (CSF). In addition, FTA-ABS turns positive earlier and remains positive longer than VDRL. Other treponemes, such as T. pertenue, may also produce a positive FTA-ABS. The ABS suffix refers particularly to a processing step used to remove nonspecific antispirochetal antibodies present in normal serum.

A latex fixation test, also called a latex agglutination assay or test, is an assay used clinically in the identification and typing of many important microorganisms. These tests use the patient's antigen-antibody immune response. This response occurs when the body detects a pathogen and forms an antibody specific to an identified antigen present on the surface of the pathogen.

<span class="mw-page-title-main">Heterophile antibody test</span> Diagnosistic test for infectious mononucleosis

The mononuclear spot test or monospot test, a form of the heterophile antibody test, is a rapid test for infectious mononucleosis due to Epstein–Barr virus (EBV). It is an improvement on the Paul–Bunnell test. The test is specific for heterophile antibodies produced by the human immune system in response to EBV infection. Commercially available test kits are 70–92% sensitive and 96–100% specific, with a lower sensitivity in the first two weeks after clinical symptoms begin.

A nontreponemal test (NTT) is a blood test for diagnosis of infection with syphilis. Nontreponemal tests are an indirect method in that they detect biomarkers that are released during cellular damage that occurs from the syphilis spirochete. In contrast, treponemal tests look for antibodies that are a direct result of the infection thus, anti-treponeme IgG, IgM and to a lesser degree IgA. Nontreponemal tests are screening tests, very rapid and relatively simple, but need to be confirmed by treponemal tests. Centers for Disease Control and Prevention (CDC)-approved standard tests include the VDRL test, the rapid plasma reagin (RPR) test, the unheated serum reagin (USR) test, and the toluidine red unheated serum test (TRUST). These have mostly replaced the first nontreponemal test, the Wassermann test.

<span class="mw-page-title-main">Hemagglutinin</span>

In molecular biology, hemagglutinins are receptor-binding membrane fusion glycoproteins produced by viruses in the Paramyxoviridae family. Hemagglutinins are responsible for binding to receptors on red blood cells to initiate viral attachment and infection. The agglutination of red cells occurs when antibodies on one cell bind to those on others, causing amorphous aggregates of clumped cells.

Virus quantification involves counting the number of viruses in a specific volume to determine the virus concentration. It is used in both research and development (R&D) in commercial and academic laboratories as well as production situations where the quantity of virus at various steps is an important variable. For example, the production of viral vaccines, recombinant proteins using viral vectors and viral antigens all require virus quantification to continually adapt and monitor the process in order to optimize production yields and respond to ever changing demands and applications. Examples of specific instances where known viruses need to be quantified include clone screening, multiplicity of infection (MOI) optimization and adaptation of methods to cell culture. This page discusses various techniques currently used to quantify viruses in liquid samples. These methods are separated into two categories, traditional vs. modern methods. Traditional methods are industry-standard methods that have been used for decades but are generally slow and labor-intensive. Modern methods are relatively new commercially available products and kits that greatly reduce quantification time. This is not meant to be an exhaustive review of all potential methods, but rather a representative cross-section of traditional methods and new, commercially available methods. While other published methods may exist for virus quantification, non-commercial methods are not discussed here.

TPPA may refer to:

<span class="mw-page-title-main">Meningeal syphilis</span> Medical condition

Meningeal syphilis is a chronic form of syphilis infection that affects the central nervous system. Treponema pallidum, a spirochate bacterium, is the main cause of syphilis, which spreads drastically throughout the body and can infect all its systems if not treated appropriately. Treponema pallidum is the main cause of the onset of meningeal syphilis and other treponemal diseases, and it consists of a cytoplasmic and outer membrane that can cause a diverse array of diseases in the central nervous system and brain.

References

  1. Cheesbrough, M. District Laboratory Practice in Tropical Countries, Part 2, Cambridge University Press, South Africa, second edition, 2006.
  2. 1 2 British Medical Journal (bestpractice.bmj.com) > Syphilis infection > Diagnostic tests Last updated: Mar 22, 2012. In turn citing:
    • Creegan L, Bauer HM, Samuel MC, et al. An evaluation of the relative sensitivities of the venereal disease research laboratory test and the Treponema Pallidum particle agglutination test among patients diagnosed with primary syphilis. Sex Transm Dis. 2007;34:1016–1108.
    • Manavi K, Young H, McMillan A. The sensitivity of syphilis assays in detecting different stages of early syphilis. Int J STD AIDS. 2006;17:768–771.
  3. Marks, M; Lebari, D; Solomon, AW; Higgins, SP (September 2015). "Yaws". International Journal of STD & AIDS. 26 (10): 696–703. doi: 10.1177/0956462414549036 . PMC   4655361 . PMID   25193248.