Nested PCR for Specific Diagnosis of
            <i>Taenia solium</i>
            Taeniasis

Mayta, Holger; Gilman, Robert H.; Prendergast, Emily; Castillo, Janeth P.; Tinoco, Yeny; Garcı́a, Héctor H.; González, Armando E.; Sterling, Charles R.

doi:10.1128/jcm.01172-07

Cited by 82 publications

(82 citation statements)

References 31 publications

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“…All of these serum samples were collected in regions where the transmission of cysticercosis/taeniasis does not occur, except for the serum samples from persons with Taenia saginata taeniasis, which were collected in Peru. Differentiation of T. solium and T. saginata was made after examining the proglottids or by PCR (18)(19)(20).…”

Section: Methodsmentioning

confidence: 99%

Recombinant Protein- and Synthetic Peptide-Based Immunoblot Test for Diagnosis of Neurocysticercosis

et al. 2014

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One of the most well-characterized tests for diagnosing neurocysticercosis (NCC) is the enzyme-linked immunoelectrotransfer blot (EITB) assay developed at the CDC, which uses lentil lectin-bound glycoproteins (LLGP) extracted from Taenia solium cysticerci. Although the test is very reliable, the purification process for the LLGP antigens has been difficult to transfer to other laboratories because of the need for expensive equipment and technical expertise. To develop a simpler assay, we previously purified and cloned the diagnostic glycoproteins in the LLGP fraction. In this study, we evaluated three representative recombinant or synthetic antigens from the LLGP fraction, individually and in different combinations, using an immunoblot assay (recombinant EITB). Using a panel of 249 confirmed NCC-positive and 401 negative blood serum samples, the sensitivity of the recombinant EITB assay was determined to be 99% and the specificity was 99% for diagnosing NCC. We also tested a panel of 239 confirmed NCC-positive serum samples in Lima, Peru, and found similar results. Overall, our data show that the performance characteristics of the recombinant EITB assay are comparable to those of the LLGP-EITB assay. This new recombinant-and synthetic antigen-based assay is sustainable and can be easily transferred to other laboratories in the United States and throughout the world.T he diagnosis of neurocysticercosis (NCC), a condition caused by the larvae Taenia solium, is reliably established using results from both imaging and serological tests. The guidelines proposed for making a diagnosis of presumptive NCC (1, 2) define cases as definitive, probable, or possible based on computed tomography (CT) or magnetic resonance imaging (MRI) findings, exposure risk, and serological results. These guidelines specifically define the lentil lectin-bound glycoprotein enzyme-linked immunoelectrotransfer blot assay (LLGP-EITB) as the serological reference standard for diagnosing NCC (3).The LLGP-EITB assay is an immunoblot method that detects antibodies to one or more of seven lentil lectin-bound glycoproteins, which are present in the soluble fraction of an extract of T. solium cysts. In patients with multiple enhancing intracranial lesions, the LLGP-EITB assay is 100% specific and 95% sensitive using blood serum or cerebrospinal fluid (CSF) samples (3-6). The original study that described and evaluated the LLGP-EITB assay was performed using serum samples from biopsy-proven cases of NCC, most with multiple lesions, as detected by skeletal radiographs, and reported a sensitivity of 98% and a specificity of 100% (3). Continued monitoring of the test performance compared with clinical findings using newer imaging techniques, such as CT and MRI, revealed that the sensitivity of the assay was lower, between 50 to 80%, in cases with a single lesion or calcified cysts (6-10). The specificity of the LLGP-EITB assay has been remarkable at essentially 100%, with only rare anecdotal reports of falsepositive results (11)(12)(13).Although the LLGP-...

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Section: Methodsmentioning

confidence: 99%

Recombinant Protein- and Synthetic Peptide-Based Immunoblot Test for Diagnosis of Neurocysticercosis

et al. 2014

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“…Such tests offer extremely high sensitivity and specificity and are often used in conjunction with magnetic resonance imaging (MRI) or computed tomography (CT) scanning to confirm the presence of Taenia cysts in neurocysticercosis. Recently, a specific nested PCR technique for the diagnosis of T. solium infection has been developed and shows promise under field conditions (Mayta et al 2008). The launch of the T. solium genomesequencing project (Aguilar-Díaz et al 2006) will aid gene discovery in this parasite and will in no doubt lead to future molecular diagnostic tests for human cysticercosis.…”

Section: Zoonotic Cestode Infections (A) Cysticercosismentioning

confidence: 99%

Zoonotic helminth infections with particular emphasis on fasciolosis and other trematodiases

Robinson

Dalton

2009

Phil. Trans. R. Soc. B

148

109

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Zoonotic infections are among the most common on earth and are responsible for .60 per cent of all human infectious diseases. Some of the most important and well-known human zoonoses are caused by worm or helminth parasites, including species of nematodes (trichinellosis), cestodes (cysticercosis, echinococcosis) and trematodes (schistosomiasis). However, along with social, epidemiological and environmental changes, together with improvements in our ability to diagnose helminth infections, several neglected parasite species are now fast-becoming recognized as important zoonotic diseases of humans, e.g. anasakiasis, several fish-borne trematodiasis and fasciolosis. In the present review, we discuss the current disease status of these primary helminth zoonotic infections with particular emphasis on their diagnosis and control. Advances in molecular biology, proteomics and the release of helminth genome-sequencing project data are revolutionizing parasitology research. The use of these powerful experimental approaches, and their potential benefits to helminth biology are also discussed in relation to the future control of helminth infections of animals and humans.

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“…The coproantigen detection method has been shown to be more sensitive than the microscopy method, but it cannot differentiate between Taenia species because of it is genus specific, not species specific (2). By contrast, various copro-DNA detection methods using PCR have been developed for sensitive differential detection of taeniid cestodes (6,12,18,25). Although these techniques provide sensitive and reliable diagnostic results, it is not easy to exploit in the laboratories of developing countries where these diseases are endemic, because PCR requires sophisticated equipment, such as a thermal cycler.…”

mentioning

confidence: 99%

“…Diagnosis is mainly performed by microscopic observation of eggs in feces and/or by comparative morphology of proglottids or scolices, but these methods lack both sensitivity and specificity. In order to overcome the lower sensitivity of microscopic diagnosis, various immunological or molecular approaches, including coproantigen and copro-DNA detection methods, have been developed (2,6,12,18,25). The coproantigen detection method has been shown to be more sensitive than the microscopy method, but it cannot differentiate between Taenia species because of it is genus specific, not species specific (2).…”

mentioning

confidence: 99%

Loop-Mediated Isothermal Amplification Method for Differentiation and Rapid Detection of Taenia Species

Nkouawa

Sako²,

Nakao³

et al. 2009

J Clin Microbiol

112

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Rapid detection and differentiation of Taenia species are required for the control and prevention of taeniasis and cysticercosis in areas where these diseases are endemic. Because of the lower sensitivity and specificity of the conventional diagnosis based on microscopical examination, molecular tools are more reliable for differential diagnosis of these diseases. In this study, we developed and evaluated a loopmediated isothermal amplification (LAMP) assay for differential diagnosis of infections with Taenia species with cathepsin L-like cysteine peptidase (clp) and cytochrome c oxidase subunit 1 (cox1) genes. LAMP with primer sets to the cox1 gene could differentiate between three species, and LAMP with primer sets to the clp gene could differentiate Taenia solium from Taenia saginata/Taenia asiatica. Restriction enzyme digestion of the LAMP products from primer set Tsag-clp allowed the differentiation of Taenia saginata from Taenia asiatica. We demonstrated the high specificity of LAMP by testing known parasite DNA samples extracted from proglottids (n ‫؍‬ 100) and cysticerci (n ‫؍‬ 68). LAMP could detect one copy of the target gene or five eggs of T. asiatica and T. saginata per gram of feces, showing sensitivity similar to that of PCR methods. Furthermore, LAMP could detect parasite DNA in all taeniid egg-positive fecal samples (n ‫؍‬ 6). Due to the rapid, simple, specific, and sensitive detection of Taenia species, the LAMP assays are valuable tools which might be easily applicable for the control and prevention of taeniasis and cysticercosis in countries where these diseases are endemic.

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Nested PCR for Specific Diagnosis of Taenia solium Taeniasis

Cited by 82 publications

References 31 publications

Recombinant Protein- and Synthetic Peptide-Based Immunoblot Test for Diagnosis of Neurocysticercosis

Recombinant Protein- and Synthetic Peptide-Based Immunoblot Test for Diagnosis of Neurocysticercosis

Zoonotic helminth infections with particular emphasis on fasciolosis and other trematodiases

Loop-Mediated Isothermal Amplification Method for Differentiation and Rapid Detection of Taenia Species

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