Table of Content

30 October 2016, Volume 34 Issue 5

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Antigenic Characterization of Immunodominant Epitopes of Alpha-8 giardin in Giardia lamblia

WEI Chao-jun1, JIA Yan-juan1, XU Hui1, WU Ling1, LU Si-qi2, LI Yong-hong1*

2016, 34(5):  1-389-393. 

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Objective To identify the dominant epitopes of α-8 giardin in Giardia lamblia, and analyze their immunoreactivity. Methods Three dominant epitopes G1（7-17 aa）, G2（30-40 aa） and G3（296-306 aa） were predicted for α-8 giardin by bioinformatic analysis using DNAstar, Biosun and CLUSTAL W softwares. Each epitope was conjugated to keyhole limpethemocyanin（KLH） and then used to immunize female Chinchilla rabbits via multi-point（8-10 points） subcutaneous injections on the back（0.5 mg conjugate/rabbit, 100 μl at each point）. Then the rabbits were boosted（0.2 mg conjugate/rabbit） on days 14, 21 and 28 after the first immunization. Blood was collected prior to each immunization, and carotid artery blood（50 ml） was collected on day 7 after the last immunization. The antibody titer for anti-KLH-G1, anti-KLH-G2 and anti-KLH-G3 sera was determined with indirect ELISA. The reactivity of the three anti-sera with recominant giardin（rGiardin） was analyzed with Western blotting. Results The concentration of the purified conjugates KLH-G1, KLH-G2, and KLH-G3 was 0.66, 0.95 and 0.25 mg/ml， respectively. ELISA showed that the antibody titer for anti-KLH-G1, anti-KLH-G2 and anti-KLH-G3 sera was 1∶12800, 1∶51200, 1∶51200, respectively. SDS-PAGE revealed a specific band at Mr 36 000 for the three anti-sera. Western blotting showed that the three anti-sera all specifically recognized rGiardin. Conclusion Immunization with the KLH-G1, KLH-G2, and KLH-G3 conjugates induced production of specific IgG antibody in rabbits, which can recognize rGiardin.

Screening and Applications of qPCR Primers for apomucin Gene of Echinococcus multilocularis

SU Meng1,2, GUO Xiao-la2, YANG Jing2, SHAO Zhong-wei2, DING Jun-tao3, XIANG Hai-tao1, LUO Xue-nong2,4, ZHENG Ya-dong2,4, SUN Xiao-lin1*

2016, 34(5):  2-394-398. 

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Objective To screen for the optimal qPCR primers for Echinococcus multilocularis apomucin gene（Em-apo） and analyze Em-apo expression. Methods Primers were designed based on 4 Em-apo sequences from GeneDB. Primer specificity and PCR efficiency were determined, based on which the optimal primer pairs were selected. Alterations of Em-apo expression in 1 000 E. multilocularis protoscoleces treated with albendazole（5 μg/ml） and insulin（100 ng/ml） were separately assessed using the selected primers. DMSO used in albendazole dilution and in PBS insulin dilution were used as the control. Results Specific primers for Em-apo-1, Em-apo-2/3, Em-apo-4 and actin were selected. qPCR melting curves revealed a single peak for each primer pair and an amplification efficiency from 95% to 101%. The qPCR showed increased expression of Em-apo-1（1.51±0.27）, Em-apo-2/3 （1.39±0.30） and Em-apo-4（1.14±0.18） after albendazole treatment in comparison to the DMSO control（1.00）（P>0.05 among the three genes）; and an unaltered Em-apo-1 expression, slightly decreased Em-apo-4 expression, and significantly decreased Em-apo-2/3 expression（0.73±0.09） after insulin treatment in comparison to the PBS control（P>0.05 among the three genes）. Conclusion The selected specific primers for Em-apo genes can be used to analyze the gene expression by qPCR. Treatment with albendazole and insulin show certain effects on the expression of Em-apo genes in E. multilocularis protoscoleces.

Analysis of Pfcrt Gene Polymorphism in Imported Falciparum Malaria Cases in Henan Province in 2015

ZHOU Rui-min1，WANG Yi-nan2，QIAN Dan1，LI Su-hua1，LIU Ying1，YANG Cheng-yun1，ZHAO Yu-ling1，XU Bian-li1，ZHANG Hong-wei1*

2016, 34(5):  3-399-404. 

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Objective To analyze Plasmodium falciparum chloroquine resistant transporter（Pfcrt） gene polymorphism in imported falciparum malaria cases in Henan Province in 2015. Methods Blood samples were collected from 132 cases of imported falciparum malaria in Henan Province in 2015. DNA was extracted from the samples, and the Pfcrt was amplified by nested PCR using specific primers. The PCR products were digested by restriction endonuclease enzyme ApolⅠ and sequenced. Pfcrt gene polymorphism and distribution were analyzed. Results Most of the 132 cases of imported malaria were young male adults returning from the Africa, with the highest percentage in those from West Africa（38.6%, 51/132）, then Central Africa（26.5%, 35/132）, South Africa（25.0%, 33/132）, East Africa（8.3%, 11/132）, and North Africa（1.5%, 2/132）. The nested PCR yielded a 145-bp product for each sample, and 66.7%（88/132） of the products were completely digested by ApolⅠ, resulting in two fragments of 114 bp and 31 bp; 32.6%（43/132） could not been digested and only a single fragment of 145 bp was shown; and 0.8%（1/132） were incompletely digested, yielding three fragments of 145 bp, 114 bp and 31 bp. By blasting against chloroquine sensitive strain 3D7, we found mutations of Pfcrt at sites correspondig to residues 74, 75 and 76 from ATG, AAT and AAA to ATT, GAA and ACA （i.e. M74I, N75E and K76T） in 43 of the 132 blood samples, and mixed type mutations into ATG/T, A/GAA/T and AA/CA at sites correspondig to residues 74, 75 and 76（CVM/I, N/E/D/K, T/K） in one blood sample. The other 88 blood samples showed a wild type with no mutation（CVMNK）. Mutations occurred mainly in cases from West Africa（41.2%, 21/51）, then East Africa（36.4%, 4/11）, South Africa（30.3%, 10/33）, and Central Africa（22.9%, 8/27）（χ2=4.07, P＞0.05）. The 2 cases from the North Africa both had wild type Pfcrt; the one with mixed type mutation was from West Africa. Conclusion Three haplotypes of Pfcrt have been found, including wild type（CVMNK）, mutation type（CVIET） and mixed type（CVM/I, N/E/D/K, K/T） in the imported malaria cases. The wild type occupies the highest proportion （66.7%）, while the mutation type possesses a high proportion of 41.2% in cases from West Africa.

Epidemiological Survey on Intestinal Parasitic Infections in Lhasa City in 2014

LIU Jian-feng1,2, JIN Xiao-lin1,2, YANG Kun1,2, XU Jin-shui3, QIAN Yi-xin1,2,YAN Wei-an1,2, ZHA Xi4, GE Sang Zhuo Ga4, YUAN Dan Wang Mu4,

2016, 34(5):  4-405-408. 

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Objective To investigate the prevalence of parasitic infections in Lhasa, Tibet. Methods The residents in Chengguan District, Dangxiong County, Dazi County and Qushui County were selected to participate in the study in 2014, using the stratified random sampling method. Infections with the soil-borne nematodes in feces were examined in triplicates using the Kato-Katz method. Further, the Enterobius vermicularis were examined using the cellophane anal swab method in children aged 3-12 years, and the intestinal protozoa were detected by iodine staining. The parasitic infection was analyzed by sex, age, and occupation. Comparisons were made using Chi-square test. Results A total of 1 015 residents（39.3±18.2 years: age range, 1-89 years） participated in the study, including 391 males（38.6%） and 623 females（61.4%）. Two hundred and forty-nine participants were positive for parasitic infections, with an overall infection rate of 24.5%（249/1 015）. The infection rate was 21.7%（220/1 015） for Taenia sp., 0.2%（2/1 015） for Trichuris trichiura and E. vermicularis respectively, 4.1%（42/1 015） for Entamoeba coli, 0.4%（4/1 015） for Giardia lamblia, 0.2%（2/1 015） for Iodamoeba butschlii. No infection was found for other parasites. There was no significant sexual difference in the infection rate of Taenia（male 21.2%, 83/391; female 22.0%, 137/624）, T. trichiura（male 0; female 0.3%, 2/624）, E. vermicularis（male 0.3%, 2/391; female 0.2%, 1/624）, E. coli（male 5.4%, 21/391; female 3.4%, 21/624）, G. lamblia（male 0.8%, 3/391; female 0.2%, 1/624）, or I. butschlii（male 0; female 0.3%, 2/624）（P>0.05）. The Taenia sp. infection rate in the age groups of 0-17, 18-29, 30-39, 40-49, 50-59 and ≥ 60 years was 10.9%（18/165）, 14.0%（18/129）, 23.0%（43/187）, 25.3%（61/241）, 26.5%（41/155）, and 28.3%（39/138）, respectively, with significant differences between groups of 0-17 and 40-49 years and between groups of 50-59 and ≥ 60 years（P<0.01）. The infection rate of E. coli, T. trichiura and I. butschlii was highest in the age group of 50-59 years（6.5%, 10/155; 0.7%, 1/155; 1.3%, 2/155 respectively）. Among various occupations, farmers showed the highest infection rate for Taenia（44.5%, 147/330）, E. coli（6.4%, 21/33）, E. vermicularis （0.6%, 2/330） and I. butschlii（0.3%, 1/330）; housewives had the highest infection rate for T. trichiura（2.5%, 1/40）; and the students had the highest infection rate for G. lamblia（0.7%, 1/134）. Conclusion There are infections with various intestinal parasites in the participants of Lhasa City, among which taenia infection reaches 20.7% while others have a low level of infection.

Epidemiological Studies on Echinococcosis in Kizilsu Kirgiz Autonomous Prefecture of Xinjiang

CHEN Xiao-ying1, Yusupjan SETIWALDI1, Yisilayin OSMAN2*

2016, 34(5):  5-409-413. 

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Objective To understand the prevalence and characteristics of echinococcosis in Kizilsu Kirgiz Autonomous Prefecture（abbreviated as Kizilsu） of Xinjiang, in the aim of providing evidence for prevention and treatment of echinococcosis. Methods Sixty-two villages were selected as investigation spots from 19 towns in agricultural and pasturing areas of Ulugqat County, Artux City, Akto County, and Akqi County during 2011-2012. Ultrasound abdominal scan was performed in all residents in these spots to examine the occurance of echinococcosis. ELISA was performed to detect the echinococcosis-specific antibody in sera from children of 6-12 years and the antigen of Echinococcus granulosus in freshly-collected dog feces. Hydatid cysts in viscera of at slaughterhouses in the four spots were examined by touching and visual observation. Results Ultrasound was conducted in 12 000 residents, among whom 60 were found to be positive for echinococcosis（echinococcosis granulosa）, with a morbidity of 0.5%（60/12 000）. The morbidity in Ulugqat County was 1.1%（35/3 200）, significantly higher than that in Artux City（0.5%, 13/2 400）, Akto County（0.2%, 8/3 200）, and Akqi County（0.1%, 4/3 200）（χ2=4.92-24.79, P<0.05）. The morbidity was highest in residents ≥71 years（1.2%, 4/323）, followed by the age group of 0-10 years（0.9%, 3/350）, but with no significant difference among age groups（χ2=0.39-3.31, P＞0.05）. The serum antibody positive rate in children was 3.1%（230/7 338）, highest in Artux City（4.9%, 89/1 828） with significant difference in comparison to Akto County（2.9%, 56/1 927）, and Akqi County（0.7%, 11/1 639）（χ2=9.71, 54.34, P<0.05）. The hydatid cyst rate in livestock was 7.7%（291/3 769）, of which 97.3%（283/291） were in sheep. The antigen positive rate in dog feces was 3.3%（74/2 219）. Conclusion The morbidity（1.1%） was highest in Ulugqat County and the antibody positive rate（4.9%） in children was highest in Artux City. Infection was found in livestock and dogs.

Serological Analysis of Antibodies against Cysticercus cellulosae and Toxoplasma gondii in 122 Patients with Meningitis Encephalitis Syndrome

SUN Lei, WANG Yan-juan*, HU Yuan, CAO Sheng-kui, SHEN Yu-juan, CAO Jian-ping

2016, 34(5):  6-414-417. 

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Objective To examine the IgG and IgM antibodies for parasites Cysticercus cellulosae and Toxoplasma gondii in 122 patients with meningitis encephalitis syndrome, and provide basis for clinical diagnosis of the meningitis encephalitis syndrome. Methods The sera were collected from patients with meningitis encephalitis syndrome in Shanghai Jiaotong University Affiliated Sixth People’s Hospital, People’s Hospital of Danyang City, and Jiangsu University Affiliated Hospital from August, 2014 to December, 2015. Serum IgG and IgM antibodies for cysticercus and T. gondii were examined using antibody test kits. The antibody positive rate was calculated and its distribution was analyzed by gender, season, age and occupation. Results A total of 122 patients with meningitis encephalitis syndrome were included. Seventeen and 22 patients of them were positive for IgG （13.9%, 17/122） and IgM（18.0%, 22/122） against cysticercus, respectively, while 29 and 8 cases were positive for IgG （23.8%, 29/122） and IgM （6.6%, 8/122） against T. gondii. The positive rate of cysticercus and T. gondii in males was 30.6%（22/72） and 31.9%（23/72） respectively, while that in females was 26.0%（13/50） and 24.0% （12/50）. The positive rate of IgM against cysticercus was 12.0%（3/25）, 27.0%（17/63）, 6.9% （2/29）, and 0（0/5） from spring to winter, highest within 13-25 years（45.0%, 9/20） among age groups, and highest in workers（7/14） among various occupations. The positive rate of IgM against T. gondii was 4.0%（1/25）, 11.1% （7/63）, 0（0/29）, and 0（0/5） from spring to winter, highest in ages >65 years（44.0%, 11/25）, and highest in patients with other occupations（4/10）. There was no statistically significant difference in the positive rate between males and females, and among different seasons, ages and occupations. Conclusion The positive rate of antibodies against cysticercus and T. gondii is high in the patients included, suggesting that a serological test for parasite infection might be performed during clinical diagnosis of meningitis encephalitis syndrome.

Risk Assessment of Schistosomiasis Transmission by Catalytic Models Based on Antibody Positive Rates

WANG Qiang, FENG Ting, QIN Zhi-qiang, ZHU Hong-qing, ZHANG Li-juan, XU Jing*

2016, 34(5):  7-418-423. 

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Objective To evaluate the transmission risk and features of schistosomiasis by analyzing the data concerning positive rates of anti-schistosome antibody using catalytic models. Methods Cross-sectional survey was conducted in seven villages with different endemicity of schistosomiasis in Jiangxi （Caohui, Xinhua, Jingtou villages）, Anhui（Yuye, Tieguai, Longshang villages） and Hubei（Hebei village） Provinces in 2008. Serum samples were collected and indirect hemagglutination assay was performed to detect anti-schistosome antibodies in serum. Antibody positive rate was calculated and differences among villages and age groups as well as between genders were analyzed. Data of antibody positive rate based on age strata for each village were analyzed by two-stage catalytic model and reversible and two-stage compound catalytic model. Parameters of each model were estimated through the maximum likelihood method. Seroconversion rate and sero-negative conversion rate were estimated in the two-stage catalytic model. Seroconversion rate, seroreversion rate and sero-negative conversion rate were estimated in reversible and two-stage compound catalytic model. The fitting effect was evaluated through correlation analysis and chi-square tests. The best fitted models and parameters were used to analyze the transmission risk and characteristics of schistosomiasis. Results A total of 6 428 individuals were examined with an average schistosomiasis antibody positive rate of 39.80%（2 485/6 428）. In terms of age, the peak of antibody positive rate in Caohui, Xinhua, Jingtou, Yuye, Tieguai, Longshang, and Hebei villages occurred in the age group of 46-50 （82.86%, 58/70）, 31-35 （60.78%, 31/51）, 31-35 （68.42%, 26/38）, 41-45（55.04%, 71/129）, 51-55 （62.38%, 63/101）, 56-60 （31.43%, 33/105）, and 16-20 （21.88%, 7/32） years old, respectively. In general, the antibody positive rate showed a trend of increase followed by a decrease with increasing age in each village. The best model for the data of Caohui, Xinhua and Jingtou and Yuye village was the two-stage catalytic model. The estimated seroconversion rate in these villages was 0.049 5, 0.044 0, 0.055 7, and 0.034 4 respectively, all higher than the corresponding sero-negative conversion rate of 0.005 9, 0.019 6, 0.015 5, and 0.017 8. The best model for the data of Longshang, Tieguai and Hebei villages was the reversible and two-stage compound model. The seroconversion rate in these villages was 0.062 9, 0.168 1, and 0.039 4 respectively, the seroreversion rate was 0.168 8, 0.121 1, and 0.152 2, and the sero-negative conversion rate was 0.001 7, 0.000 2, and 0.090 9. Conclusions The catalytic model based on antibody positive rate by age strata could reflect the transmission rate and risk quantitatively and may provide guidance for making control strategies.

Digital Description and Identification of 11 Kinds of Principal Parasite Eggs

SHEN Hai-mo, AI Lin, CAI Yu-chun, LU Yan, CHEN Shao-hong*

2016, 34(5):  8-424-429. 

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Objective To facilitate the identification of parasite eggs using computer technology, establish the automation-based applications, and propose an algorithm for egg classification. Methods Eggs of 11 parasites, Clonorchis sinensis, Taenia solium, Enterobius vermicularis, Ascaris lumbricoides, Trichuris trichiura, Spirometra mansoni, Diphyllobothrium latum, Ancylostoma duodenale, Schistosoma japonicum, Paragonimus westermani and Fasciolopsis buski, were selected and divided into two groups, the training group and the testing group, and were microphotographed. The eigenvalue was extracted using the VC++-based method. The eigenvalue database was constructed, and the training data set was tested with a variety of classification algorithms. The classifier was constructed using algorithm with the highest efficiency and an identification method was established by multi-feature fusion. Results After removal of images with invalid values, the training group received 19 844 egg images, and the testing group, 3 721 images. Based on the 14 eigenvalues, there were significant differences in the size and color among the eggs of 11 parasite species. For example, the length, width, area and brightness of the smallest parasite egg of Clonorchis sinensis were 292.24 μm, 192.64 μm, 43 416.61 μm2 and 53.84, respectively, while those of the largest parasite egg of Fasciolopsis buski were 945.31 μm, 610.88 μm, 536 002.60 μm2 and 100.54, respectively. When using dynamic weights to construct the classifier, the discrimination rate on the training data set was 88.89%（17 641/19 844）, and that on the verification data set was 91.83%（3 004/3 271）, with an average modeling time of 0.01 s. Conclusion The algorithm for egg classification has been established, which pravides a basis for further study on its feasibility．

Evaluation on Malaria Hotspots in Yingjiang County of the China-Myanmar Border Area in 2015

ZHANG Cang-lin1, GUO Xiang-rui2, YANG Rui1, YANG Ya-ming1, YANG Zhong-hua1,ZHENG Yu-ting1, WU Chao1, LI Shi-gang3, LIN Ying-xue2, YU Guo-cui1, YAN Run-xian2,

2016, 34(5):  9-430-434. 

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Objective To understand the control status of malaria at hotspots in Yingjiang County and provide measures for malaria elimination in the China-Myanmar border areas of Yunnan Province. Methods A survey was made in 4 villages with indigenous malaria cases or imported cases in Nabang and Tongbiguan of Yingjiang County in Yunnan Province in June and July 2015. Peripheral blood samples were collected from the neighboring residents around patients and examined by malaria rapid diagnostic test （RDT）. The results were further verified by nested-PCR. Mosquitoes were collected by overnight trapping with light traps in Jingpo, Lilisu, Jiema, and Mengxiangyang villages or by human landing catches in Jingpo and Lisu villages. Nested-PCR was performed on part of the captured Anopheles minimus to detect the malaria parasites. Results One hundred and ninety-four filter blood samples were collected from 11 malaria cases in two sites. All were detected to be negative for Plasmodium by RDT. In contrast, two samples originated from Jingpo and Lisu villages with indigenous cases were detected to be positive for Plasmodium vivax by nested-PCR. A total of 2 374 mosquitoes were captured, belonging to 22 species of 4 genera: Anopheles, Culex, Aedes and Armigeres. The mosquitoes were predominated by genus Culex, followed by genus Anopheles（11.33%, 269/2 374） which was dominated by A. minimus（49.07%, 132/269）, then was A. sinensis（4.09%, 11/269）, A. maculatus（2.23%, 6/269）, A. jeyporiensis（0.74%, 2/269）and so on. The mean indoor man-biting rate of mosquitoes was 5.78 and 3.20 per person per hour for Jingpo and Lisu villages, and the mean outdoor man-biting rate of mosquitoes was 2.30 per person per hour for Lisu Village. The 14 A. minimus were negative for sporozoite infection as detected by nested-PCR. Conclusion Nested-PCR showed that there are asymptomatic Plasmodium carriers in Yingjiang’s border area of Yunnan Province. Four major mosquito species as malaria vectors exist with A. minimus as the dominant one.

Evaluation of Measures and Achievements of Malaria Control in Tengchong City, Yunnan Province during 2010-2015

WANG Jia-zhi1, YIN Shou-qin1, LI Sheng-guo1, LI Xi-shang1, CAI Wen-bin1, FENG Jun2*

2016, 34(5):  10-435-438. 

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Objective To evaluate the measures and achievements of malaria control in Tengchong City during 2010-2015. Methods The malaria control information on epidemiology, foci disposal, blood detection of febrile patients, and medical treatment during 2010-2015 in Tengchong City was collected and analyzed using Microsoft Excel 2010. Results In 2010-2015, 1 654 malaria cases were reported in Tengchong City, including 18 indigenous cases, 22 domestically mobile cases, and 1 614 imported cases from abroad, of whom 1 584 cases （98.1%） were imported from Myanmar. Most of the cases were vivax malaria（76.2%, 1 261/1 654）. No indigenous malaria cases were reported from 2013 to 2015. Blood test was conducted for 80 655 febrile patients, with a positive detection rate of 2.1%（1 654/80 655）. The positive detection rate was highest in 2010 （2.8%, 700/24 861）, lowest in 2011（1.4%, 341/23 623）, and decreased from 2012 to 2015. In addition, 1 654 cases were directly reported through online system. The 24-h case report rate during 2013-2015 was 100%. A total of 1 191 cases were investigated. The 3-day case investigation rate during 2013-2015 was 100%. A total of 1 351 endemic foci were investigated. The 7-day foci disposal rate during 2014-2015 was 100%. Conclusion No indigenous transmission has been reported for three years in Tengchong City. However, the imported malaria remains an important problem.

Analysis on Malaria Situation in Shaanxi Province during 2005-2014

ZHOU Ti-cao1,2,3, ZHANG Yi1, ZHANG Zhi-cheng1, DENG Yong1, CHAI Zi-chao1, CAO Lei1, CHEN Sa1, SUN Jun-ling2, SHI Guo-qing3, LIU Feng1*, LI Zhong-jie2

2016, 34(5):  11-439-443. 

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Objective To analyze the malaria situation in Shaanxi Province during 2005-2014, in the aim of providing reference for improving malaria elimination strategies and measures. Methods Data on malaria control and malaria epidemic reports during 2005-2014 were collected, and analysis was made on prevalence and regional distribution of malaria, species of plasmodium, diagnosis of patients, and sources of imported malaria. Results Four hundred and fifty three cases of malaria were reported in Shaanxi Province during 2005-2014, consisting of 73 local cases （16.1%） and 380 imported cases （83.9%） of which 141 were falciparum malaria （37.1%, 141/380）. Three patients died and they all were imported cases of falciparum malaria. No indigenous infection had been reported since 2011. Most of the cases were laboratory-confirmed（71.5%, 324/453）. The falciparum malaria cases accounted for 31.1%（141/453） of all the cases, and they all were imported cases. No cases of quartan malaria and ovale malaria were reported. The 73 indigenous cases distributed in 25 districts（counties） of 7 cities, including 3 cities in South Shaanxi （Shangluo, Hanzhong, Ankang）（61.6%, 45/73） and four cities in Central Shaanxi（Xi′an, Weinan, Xianyang, Baoji） （38.4%, 28/73）. The imported cases showed an increasing trend from 24 in 2005 to 59 in 2014. The 380 imported cases were mainly from over 20 countries in Africa （72.6%, 276/380）, with top sources of Angola （64 cases）, Cameroon （26）, Ghana （24）, and Equatorial Guinea （23） accounting for 36.0% （ 137/380）. The median of interval from onset to diagnosis was 5 d. The cases were mainly reported by clinical medical institutions （87.6%, 397/453）, only 7.5% （34/453） were reported by disease control institutions. Conclusion No indigenous cases have been reported since 2011, but the imported malaria cases show a trend of increase with a major source of Africa.

Establishment of LAMP Detection for Toxoplasma gondii Based on the 529 bp Repeat Sequence

XU Qian-ming1,2*, YUAN Heng-qing1, ZHAO Chang-cheng1, FANG Fen1, LIU Zhe1, NING Hong-rui1,2, SHI Da-li1, LI Yu1, WANG Hai-yang1

2016, 34(5):  12-444-450. 

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Objective To establish the loop mediated isothermal amplification（LAMP） technique for determining Toxoplama gondii. Methods The primers for LAMP of the conserved 529 bp sequence was designed by the Primer Explorer 4.0 software. The LAMP reaction was made on the constructed pMD-19T-529 bp recombinant plasmid as a template, and was optimized in loop primer, concentrations of betaine and MgSO4, and reaction temperature. The optimized LAMP and PCR were performed in ultra-pure water, on pig genome, Cryptosporidium parvum genome, Isospora suis genome, and pMD-19T-529 bp, respectively, to test the sensitivity of LAMP. The pMD-19T-529 bp was serially diluted to 109-100 copies/ml to test the specificity of LAMP. Results LAMP using the designed primers amplified the 529 bp fragment of T. gondii. The optimized LAMP system had a total reaction volume of 25 μl, with the optimal concentrations of betaine and MgSO4 being 0.4 mol/L and 6 mmol/L, respectively. The amplification efficiency of 30 min reaction in the presence of loop primer was comparable to that of 60 min reaction without loop primer, which indicated that addition of loop primer shortened the reaction time by 30 min. The optimal reaction condition was 63 ℃ 30 min, 80 ℃ 3 min. The established LAMP method specifically amplifed the 529 bp fragment of T. gondii, and its efficiency was 10 times of PCR（detection threshold 103 copies/ml vs. 104 copies/ml）. Conclusion The established LAMP for detecting Toxoplama gondii 529 bp repeat sequence shows high specificity and sensitivity.

Data Integration and Information Sharing of Parasite Resources in Fujian Province

LI Li-sha*, ZHANG Rong-yan, CHEN Bao-jian, LIN Chen-xin, LI Yan-rong, CHEN Zhu-yun, FANG Yan-yan, CHENG You-zhu, LIN Jin-xiang

2016, 34(5):  13-451-454. 

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Objective To establish parasite library and database in Fujian Province by integrating information on human-animal coinfecting parasites, vectors and　intermediates, in the aim of resource sharing. Methods According to the Standards and Technical Regulation for Preservation of Parasitic Germplasm Resources set by the Resource Sharing Platform for Parasitic and Tropical Diseases(referred to as Germplasm Resource Sharing Platform）,the preserved and newly discovered parasitic germplasm resources in Fujian Center for Disease Control and Prevention were categorized, information described, preserved and displayed, and the information sharing was made through the Germplasm Resource Sharing Platform. Results The library and database on parasites, medical vectors, and hosts were established. The library was comprised of 1 020 parasite specimens, vectors and hosts, including 100 （9.8%） adult worm specimens, 120（11.7%） eggs and larvae, and 800（78.4%） vectors and hosts. Information digitalization was achieved for 61 resources, including 9 species of nematodes, 16 species of trematodes, 7 species of tapeworms, 16 different medical mollusks, and 13 different medical arthropods, resulting in 3 150 pieces of information and 6 300 images. The database was integrated into the Germplasm Resource Sharing Platform for resource sharing. Conclusion The library and database on parasite gemplasm resources in Fujian Province has been established. Information sharing is made available through the Germplasm Resource Sharing Platform.

Human Demodicosis: Emerging Dermatosis Caused by Demodex

ZHAO Ya-e

2016, 34(5):  14-455-462. 

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Demodex mites were discovered 175 years ago. However, the pathogenicity of Demodex remains questioned by clinicians due to the high infection yet low incidence of demodicosis and incomplete correlation between mite loads and clinical symptoms. In the recent decades, the pathogenicity of Demodex has attracted public concern as there are continuous case reports of skin lesions caused by mite infection. The pathogenicity of Demodex remains poorly understood. Demodex infection is suspected to be assocaited with rosacea, but its relation with other types of facial lesions such as papule and pustule is usually misdiagnosed or missed in clinical practice. This review gives an overview on the discovery, typical cases, pathogenesis, and clinical diagnosis and treatment of demodicosis, in the aim of promoting social recognition and attention on demodicosis.

Research Advances on Gliding-associated Proteins of Toxoplasma gondii

LI Run-hua1, YIN Guo-rong2*

2016, 34(5):  15-463-467. 

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The ability to invade host cells is a key to the survival and pathogenicity of Apicomplexan parasites. Toxoplasma gondii is an obligatory intracellular parasite. Its motility, invasion into, and egression from host cells are powered by a machinery called acto-myosin motor （AMM）. The AMM is composed of myosin A, a myosin light chain （MLC1）, two essential light chains （ELC）1, 2 and gliding-associated protein （GAP）. The GAP family has been discovered to include GAP45, GAP50, GAP80, GAP70 and GAP40, which are the major components of glideosome that provides power for parasite motility. The glideosome of apicomplexan parasites is an actin- and myosin-based power machine located at the pellicle between the plasma membrane （PM） and inner membrane complex（IMC）. This review outlines our current understanding of GAP function and architecture as well as the molecular basis for parasite motility. Meanwhile, the use of GAPs as the candidate toxoplasmosis vaccine is prospected.

Research Progress on Risk Assessment of Secondary Transmission of Imported Malaria

LEI Lei1,2, Mamatjan UMAR1,3, LI Zhi-hong2, XIA Zhi-gui1, XIA Shang1, GONG Yan-feng2, XIAO Ning1*

2016, 34(5):  16-468. 

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In 2015, WHO issued the Global Technical Strategy for Malaria 2016-2030, which sets the target of reducing global malaria incidence and mortality rates by at least 90% by 2030. Although many countries have successfully achieved malaria elimination, they are facing the risk of imported malaria. In China, despite the acceleration of malaria elimination, imported malaria has become a potential threat to achieving complete malaria elimination. This paper reviews the worldwide research progress on risk assessment of secondary transmission of imported malaria, in the aim of providing reference for risk assessment of imported malaria and preventing secondary transmission in China.

An Investigation on a Case of Family-clustered Paragonimiasis in Wenzhou City

NI Qing-xiang1*，LIN Xiao-yao2，YAO Li-nong3，RUAN Wei3，BAO Chang-xian2，CHEN Yi1，PAN Qiong-jiao1

2016, 34(5):  17-448. 

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Paragonimus infection was reported in a family of 8 members after consuming wine-preserved raw crabs. Seven members developed symptoms of fever, cough, sputum, fatigueness, chest pain, and abdominal pain during 2-3 months after crab feeding, while one member was normal. Serum samples were collected from the 7 members for anti-paragonimus antibody test, with 5 members showing positive, 1 weakly positive and 1 negative. In view of the epidemiological history, clinical symptoms, imaging and laboratory test results, this case was diagnosed to be a family-clustered paragonimus infection.

Exploration and Application of Discussion-based Teaching Method in Teaching of Medical Parasitology

SU Ju-xiang*, CAI Lian-shun, CHEN Guang, DAI Yue, ZHANG Hui-ming,GONG Zi-lin, CHE Shi-wei

2016, 34(5):  18-473-475. 

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In this study, students majoring in Clinical Medicine were enrolled to explore the effect of discussion-based teaching method in the teaching of medical parasitology. One hundred and fifty-six students（with an entry year of 2011） in classes 1-3 received the discussion-based teaching while 153 students in classes 4-6 received traditional teaching. The effect of teaching was evaluated in terms of final examination score and questionnaire, and compared between the groups. The final examination score of students receiving the discussion-based teaching （86.1±6.6） was significantly higher than those receiving the traditional teaching（74.2±8.3）（P<0.05）. The discussion-based teaching method was graded as “excellent” by 89.1%（136/156）of the students, and was considered to be superior to the traditional teaching by 96.8%（151/156）of the students. The results indicate that the discussion-based teaching method can enhance interactions between participants, change the ways of thinking, and provide inspirations for learning and exploration.