Department of Vector
Control, National Institute of Parasitic Disease, Chinese Center for
Disease Control and Prevention, Key Laboratory of Parasite and Vector
Biology of the Ministry of Health, Shanghai 200025, China;
Abstract：Pooling field specimens could reduce the number of assay and thus increase the efficiency in detecting and screening pathogen infections by polymerase chain reaction (PCR)-based assay. We investigated a pooling strategy in diagnosis of Angiostrongylus cantonensis in Pomacea canaliculata. Two settings of specimens were prepared, divided into portions and detected by multiplex PCR. Specimens A was 0.4490 g positive lung tissue of 28 larval nodes from four snails mixed with 1.310€g negative lung tissue from six snails and divided into 32 portions. Specimens B was 0.5448 g positive lung tissue with 26 larval nodes from two snails mixed with 1.092 g negative lung tissue from seven snails and divided into 48 portions. Repeated sampling was performed and sample size-accumulated positive rate curves were drawn. According to the sample size-accumulated positive rate curves, the appropriate sample size of the two specimens were 18 and 15, respectively, which is 0.36–0.58 to the total sample size. These test characteristics and the relevant factors to the sample size would need to be determined in much larger studies and more appropriately in field populations. The result indicates that the number of larval node is not the most important, nor the only factor to the sample size. It also implies the feasibility to detect A.€cantonensis in P. canaliculata by pooling strategies.
Lv S, Zhou X N, Zhang Y, Wang X H, Yang K. Epidemiological survey ofangiostrongyliasis in mainland China. Zhongguo Dongwuxue Zazhi, 2006, 22(10): 987–988 (in Chinese)
Lv S, Zhang Y, Steinmann P, Zhou X N. Emerging angiostrongyliasis in Mainland China. Emerg Infect Dis, 2008, 14(1): 161–164 doi: 10.3201/eid1401.061529
Lv S, Zhang Y, Chen S R, Wang L B, Fang W, Chen F, Jiang J Y, Li Y L, Du Z W, Zhou X N. Human angiostrongyliasisoutbreak in Dali, China. PLoS Negl Trop Dis, 2009, 3(9): e520–e527 doi: 10.1371/journal.pntd.0000520
He H J, Wu Z D, Yu X B. Pathogenic mechanism of Angiostrongylus cantonensis. Guoji Yixue Jishengchongbing Zazhi, 2007, 34(5): 242–244 (in Chinese)
Ali A B, Enden E V D, Gompel A V, Esbroeck M V. Eosinophilic meningitis due to Angiostrongylus cantonensis in a Belgian traveler. Travel Med Infect Dis, 2008, 6(1―2): 41–44 doi: 10.1016/j.tmaid.2007.09.001
Lv S, Zhang Y, Liu H X, Zhang C W, Steinmann P, Zhou X N, Utzinger J. Angiostrongylus cantonensis: morphological and behavioral investigation within the freshwater snail Pomacea canaliculata. Parasitol Res, 2009, 104(6): 1351–1359 doi: 10.1007/s00436-009-1334-z
Lv S, Zhang Y, Liu H X, Hu L, Yang K, Sternmann P, Chen Z, Wang L Y, Utzinger J, Zhou X N. Invasive snails and an emerginginfectious disease: results from the first national survey on Angiostrongylus cantonensis in China. PLoS Negl Trop Dis, 2009, 3(2): 1–8 doi: 10.1371/journal.pntd.0000368
Lin J X, Jie H Y, Li L S. Enlightenment of angiostrongyliasis outbreaks. Zhongguo Jishengchongxue Yu Jishengchongbing Zazhi, 2005, 23(5): 341–343 (in Chinese)
Wang Q P, Lai D H, Zhu X Q, Chen X G, Lun Z R. Human angiostrongyliasis. Lancet Infect Dis, 2008, 8(10): 621–630 doi: 10.1016/S1473-3099(08)70229-9
Lv S, Zhang Y, Wang X H, Liu H X, Zhu D, Yin W G, Zhou X N. Experimental study on compatibility of three species of freshwater snails with Angistrongylus cantonensis. Zhongguo Jishengchongxue Yu Jishengchongbing Zazhi, 2006, 24(4): 277–280 (in Chinese)
Liu H X, Zhang Y, Zhou X N, Lv S, Zhu D, Lin J X, Li L S. LI Y S. Studies on the growth-development andinfectivity of Angiostrongylus cantonensis in dormant Pomacea canaliculata. Zhongguo Jishengchongxue Yu JishengchongbingZazhi, 2006, 24(4): 269–272 (in Chinese)
Liu H X. Zhang Yi, Lv S, Zhu D, Wang X H, Hu L, Zhou X N. A comparative study of threemethods in detecting Angiostrongylus cantonensis larvae in lung tissue of Pomacea canaliculata. Zhongguo Jishengchongxue Yu Jishengchongbing Zazhi, 2007, 25(1): 53–56 (in Chinese)
Zhang Y, Zhou X N, Liu H X, Lv S, Li L S, Lin J X, Li Y S. Development of PCR assay for detection of Angiostrongylus cantonensis in Pomacea canaliculata. Zhongguo Jishengchongxue Yu Jishengchongbing Zazhi, 2006, 24(5): 353–355 (in Chinese)
Wei J L, Zhou W C, Shao B Y, She S S, Wang S K, Chen W B, Chen S L. Establishment of a PCR assay for detection of snails infected with Angiostrongylus cantonensis. Zhonguo Dongwuxue Zazhi, 2008, 24(12): 1136–1140 (in Chinese)
Qvarnstrom Y, Sullivan J J, Bishop H S, Hollingsworth R, da Silva A J. PCR-based detection of Angiostrongylus cantonensis in tissue and mucus secretions from molluscan hosts. Appl Environ Microbiol, 2007, 73(5): 1415–1419 doi: 10.1128/AEM.01968-06
Yin S W, Yan H M, Wang H Q, Xu F. Study on the biology of Ampullaria gigas. Acta Sci Nat Univ Norm Hunan, 2000, 23(2): 76–82
Caldeira R L, Jannotti-Passos L K, Lira P M, Carvalho O S. Diagnostic of Biomphalaria snails and Schistosoma mansoni: DNA obtained from traces of shell organic materials. Mem Inst Oswaldo Cruz, 2004, 99(5): 499–502 doi: 10.1590/S0074-02762004000500007
Cucher M A, Carnevale S, Prepelitchi L, Labbé J H, Wisnivesky-Colli C. PCR diagnosis of Fasciola hepatica in field-collected Lymnaea columella and Lymnaea viatrix snails. Vet Parasitol, 2006, 137(1―2): 74–82 doi: 10.1016/j.vetpar.2005.12.013
Velusamy R, Singh B P, Raina O K. Detection of Fasciola gigantica infection in snails by polymerase chainreaction. Vet Parasitol, 2004, 120(1―2): 85–90 doi: 10.1016/j.vetpar.2003.11.009
Hanelt B, Adema C M, Mansour M H, Loker E S. Detection of Schistosoma mansoni in Biomphalaria using nested PCR. J Parasitol, 1997, 83(3): 387–394 doi: 10.2307/3284399
Magalhães K G, Passos L K, Carvalho Odos S. Detection of Lymnaea columella infection by Fasciola hepatica through Multiplex-PCR. Mem Inst Oswaldo Cruz, 2004, 99(4): 421–424
Hertel J, Haberl B, Hamburger J, Haas W. Description of a tandem repeated DNA sequence of Echinostoma caproni and methods for its detectionin snail and plankton samples. Parasitology, 2003, 126(Pt 5): 443–449 doi: 10.1017/S0031182003003093
Shubkin C D, White M W, Abrahamsen M S, Rognlie M C, Knapp S E. A nucleic acid-based test for detection of Fasciola hepatica. J Parasitol, 1992, 78(5): 817–821 doi: 10.2307/3283311
Liu H X, Zhang Y, Zhou X N, Lv S, Zhu D, Lin J X, Li L S, Li Y S, Yin W G. Studies on susceptibility of Pomacea canaliculata of different developmental stages to infection with Angiostrongylus cantonensis. Zhongguo Jishengchongxue Yu Jishengchongbing Zazhi, 2005, 23(5): 262–265 (in Chinese)
Ni Y, Cui D Y, Zhang Z J, Peng W X. Study on method of determination of appropriate sample size of Oncomelania hupensis in marshland and lakeregions. Zhongguo Jishengchongxue Yu JishengchongbingZazhi, 2007, 19(4): 281–284 (in Chinese)
