Serological Diagnosis of North American Paragonimiasis by Western Blot Using Paragonimus kellicotti Adult Worm Antigen

Abstract

Abstract. We studied the value of an IgG Western blot (WB) with Paragonimus kellicotti (Pk) antigen for diagnosis of North American paragonimiasis. The test was evaluated with sera from patients with Pk and Paragonimus westermani infections, with control sera from patients with other helminth infections, and sera from healthy Americans. All 11 proven Pk infection sera and two samples from suspected cases that were negative by P. westermani WB at the Centers for Disease Control and Prevention (CDC) contained antibodies to antigens at 34 kDa and at 21/23 kDa. Seven of 7 P. westermani sera contained antibodies to the 34 kDa antigen, but only 2 recognized the 21/23 kDa doublet. No control samples were reactive with these antigens. Antibody reactivity declined after praziquantel treatment. Thus, the P. kellicotti WB appears to be superior to P. westermani WB for diagnosing Pk infections, and it may be useful for assessing responses to treatment.

Systems Biology Studies of Adult Paragonimus Lung Flukes Facilitate the Identification of Immunodominant Parasite Antigens

“…Patient sera were tested for reactivity against adult P. kellicotti and P. westermani antigen by Western blot as previously described [10] . Serum samples from five strongly-reactive patients were pooled (total volume 3 mL), and total IgG was precipitated using saturated ammonium sulfate (Thermo Fisher Scientific, Pittsburg, PA), re-suspended in 1× phosphate buffered saline (PBS), and desalted by dialysis against 4L 1× PBS for 2 hours at room temperature, against 4L 1× PBS 2 hours at 4°C, and against 4L 1× PBS overnight at 4°C.…”

“…Egg detection is also insensitive due to temporal inconsistencies and requires knowledge and expertise that are not readily available in many clinical settings. Serological tests for P. westermani and P. kellicotti using native parasite antigens have been described, but these tests are impractical for widespread use because they require continued access to adult parasites [8] , [10] , [11] . Thus far, efforts to develop and implement practical, standardized molecular diagnostic tools have been hindered by a lack of information on the basic biology and genomics of Paragonimus species.…”

Surveillance and diagnosis of zoonotic foodborne parasites

“…is the detection of at least one parasite egg in any sample by three different methods, including wet film (WF), Ziehl‐Neelsen stain (ZNS), and a formalin‐ether concentration technique (FECT). A western blot assay has been designed to detect P. kellicotti and P. westermani infections for the serologic diagnosis of paragonimiasis (Fischer et al., ). This method depends on the identification of the ES Ags of the parasite.…”

Paragonimiasis

“…This approach has been used for diagnosis of paragonimiasis by the Centers for Disease Control, Atlanta, Georgia in the USA, since 1988, the antigen generally used being a crude extract from P. westermani . However, extracts from P. kellicotti are better for diagnosis of that species [ 163 ].…”

“…Nevertheless, work to produce draft genomes of several Paragonimus species has now started ( http://genome.wustl.edu/projects/detail/ helminth-sequencing-project/ ). Transcriptome data for P. kellicotti produced as part of this sequencing project was used to show that genes encoding diagnostic antigens in P. westermani differ substantially from their orthologs in P. kellicotti [ 163 ].…”