Objective: To improve the sensitivity, specificity and stability of the Tag-primer nested/multiplex PCR for malaria diagnosis.
Methods: Filter paper blood samples were collected from 30 non-malaria fever patients and 20 infectious disease patients (common cold, influenza, typhoid, hepatitis, etc.). Four ml blood each taken from one falciparum malaria patient and one vivax malaria patient was serially diluted. Healthy blood sample was used as negative control. Improved direct heating method was used to prepare DNA template. The cytochrome oxidase gene (coxI) located in mitochondrion was selected as target gene. Relevant web resources and software (PUBMED, NCBI-BLAST, Mfold server and Primer Premier 5.0) were employed to design and optimize Tag-primer nested/multiplex PCR (UT-PCR) which was used to test all blood samples.
Results: A 611 bp band and a 255 bp band were seen in serially diluted infected blood samples (1,000, 100, 10 and 1 parasite/microl) from P.f and P.v patient tested by UT-PCR. The detection limit of either P. falciparum or P. vivax reached 1 parasite/microl, and the tested blood samples of non-malaria fever patients, patients with other infectious diseases and healthy persons were all negative. Consistent results of each sample in more than 3 duplicated tests were obtained.
Conclusion: The optimized Tag-primer nested/multiplex PCR shows high sensitivity, specificity and stability in malaria diagnosis.
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Verification of clinically diagnosed cases during malaria elimination programme in Guizhou Province of China.
Malar J
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Background: China is implementing a National Malaria Elimination Programme. A high proportion of clinically diagnosed malaria cases is reported in some provinces of China. In order to understand the exact situation and make clear the nature of these patients, it is of much importance to make case verifications, particularly from the pathogenic perspective.
View Article and Find Full Text PDF[Primary evaluation on the application of nested/multiplex PCR in malaria diagnosis and surveillance].
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi
August 2008
Objective: To compare the usefulness of Tag-primer nested/multiplex PCR (UT-PCR) method with microscopy in malaria diagnosis and surveillance.
Methods: 400 blood smears and blood samples on filter paper were taken from febrile patients which were initially diagnosed as malaria or suspected malaria during surveillance in mixed endemic areas of Plasmodium falciparum (Pf) and P. vivax (Pv) in Hainan and Yunnan provinces and a malaria-controlled area in Guangxi Zhuang Autonomous Region.
[Sensitivity, specificity and stability of the Tag-primer nested/multiplex PCR for malaria diagnosis].
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi
June 2007
Guangxi Center for Disease Control and Prevention, Nanning 530021, China.
Objective: To improve the sensitivity, specificity and stability of the Tag-primer nested/multiplex PCR for malaria diagnosis.
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[Tag primer-nested/ multiplex PCR for detection of Plasmodium falciparum and Plasmodium vivax].
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi
June 2005
Guizhou Provincial Center for Diseases Control and Prevention, Guiyang 550004, China.
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