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Clinical and Vaccine Immunology, July 2009, p. 1047-1051, Vol. 16, No. 7
1071-412X/09/$08.00+0     doi:10.1128/CVI.00113-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Hemagglutination Test for Rapid Serodiagnosis of Human Pythiosis{triangledown}

Thanyasiri Jindayok,1 Savittree Piromsontikorn,1 Somboon Srimuang,2 Kalayanee Khupulsup,1 and Theerapong Krajaejun1*

Clinical Immunology Laboratory, Department of Pathology,1 Research Center, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Bangkok, Thailand2

Received 12 March 2009/ Returned for modification 2 April 2009/ Accepted 26 May 2009

Human pythiosis is an emerging, life-threatening infectious disease, caused by the oomycete Pythium insidiosum. Thailand is an area where human pythiosis is endemic and the genetic blood disorder thalassemia is a predisposing factor. Patients with pythiosis present with arterial occlusions of the lower extremities, corneal ulcers, or chronic cutaneous infections. Diagnosis relies on time-consuming, relatively insensitive tests such as culture identification and immunodiffusion assay. Most patients undergo surgical removal of infected organs, and many die from the infection. Delayed diagnosis results in a poor prognosis. Here, we describe a hemagglutination (HA) test for rapid diagnosis of human pythiosis. Sheep red blood cells were coated with P. insidiosum protein extract and used in duplicated detection assays using serum samples from 33 patients with vascular (n = 27), cutaneous (n = 2), or ocular (n = 4) pythiosis and serum samples from 289 control patients with other infectious diseases (n = 77), with highly positive antinuclear antibody (n = 5), with thalassemia (n = 21), or with no known disorder (i.e., healthy blood donors) (n = 186). Based on receiver-operating characteristic analysis, a serum titer of 1:160 was selected as the cutoff point for the HA test. Serum samples that generated HA at the cutoff titer were read as positive, while samples that did not were read as negative. Positive results were obtained with the serum samples of all patients with vascular and cutaneous pythiosis and with two serum samples from the control group. Negative results were obtained with serum samples from all ocular pythiosis patients and the 287 remaining serum samples from the control group. Sensitivity and specificity of the HA were 88% and 99%, respectively. In conclusion, the HA test for detection of anti-Pythium antibodies is a simple, rapid, and reliable test for serodiagnosis of vascular and cutaneous pythiosis.

* Corresponding author. Mailing address: Department of Pathology, Faculty of Medicine, Ramathibodi Hospital, Mahidol University, Rama 6 Road, Bangkok 10400, Thailand. Phone: 662-201-1379. Fax: 662-201-1611. E-mail: mr_en{at}hotmail.com

{triangledown} Published ahead of print on 3 June 2009.

Clinical and Vaccine Immunology, July 2009, p. 1047-1051, Vol. 16, No. 7
1071-412X/09/$08.00+0     doi:10.1128/CVI.00113-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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