CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES >
Efficiency evaluation of three assays for detection of Schistosoma japonicum infections in wild mice
Received date: 2025-02-07
Revised date: 2025-03-19
Online published: 2025-04-27
Objective To evaluate the performance of liver homogenate smear microscopy (parasitological detection), real-time quantitative reverse transcription PCR (qPCR) assay and loop-mediated isothermal amplification (LAMP) for detection of Schistosoma japonicum infections in wild rodents, so as to provide an optimal laboratory diagnostic assay for surveillance of S. japonicum infection in wild rodents. Methods A total of 115 wild rodents liver samples (63 samples from Village A and 52 samples from Village B) were collected from Dongzhi County, Anhui Province, and detected by liver homogenate smear microscopy, qPCR assay and LAMP, respectively. The positive rates of the samples were calculated and compared. The consistency between qPCR and LAMP for detection of the liver samples was evaluated with microscopy as a standard, and 70% of liver samples with a low consistency between the parasitological assay and nucleic acid tests were randomly selected for validation with DNA sequencing. Results Liver homogenate smear microscopy detected 54 positive liver samples, with a positive rate of 46.96% (54/115), and the positive rate of liver samples was 46.03% (29/63) in Village A and 48.08% (25/52) in Village B (χ2 = 0.001, P > 0.05). qPCR assay tested a 69.57% (80/115) positive rate of liver samples, which was higher than liver homogenate smear microscopy (χ2 = 11.175, P < 0.01), and there positive rate was 66.67% (42/63) in Village A and 73.08% (38/52) in Village B (χ2 = 0.340, P > 0.05). LAMP tested a 56.52% (65/115) positive rate of liver samples, which was comparable to liver homogenate smear microscopy (χ2 = 1.741, P > 0.05), and the positive rate was higher in Village A(71.43%, 45/63) than in Village B (38.46%, 20/52) (χ2 = 11.293, P < 0.01). There was a high consistency between qPCR assay and liver homogenate smear microscopy (Kappa value = 0.524 4), and a low consistency between LAMP and liver homogenate smear microscopy (Kappa value = 0.154 5). Among the 24 liver samples from Village A that were negative for liver homogenate smear microscopy but positive for LAMP, 17 samples were randomly selected for DNA sequencing, and 15 of these samples showed a sequence identity of 98% and higher with the 28S ribosomal DNA of S. japonicum (GenBank accession No.: JF721395.1) and were therefore identified as positives. Conclusion Three assays have diverse sensitivities for detection of S. japonicum infections in wild rodents. qPCR assay has the highest positive rate for detection of S. japonicum infections in wild rodents and a high consistency with liver homogenate smear microscopy, while LAMP is convenient to perform but has a low consistency with liver homogenate smear microscopy.
TANG Qi , LV Chao , WANG Xi , GUO Suying , XU Xiaojuan , ZHU Hai , LI Yinlong , LIN Weina , ZHOU Xinjie , FENG Ting , XU Jing , QIN Zhiqiang . Efficiency evaluation of three assays for detection of Schistosoma japonicum infections in wild mice[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2025 , 43(2) : 186 -191 . DOI: 10.12140/j.issn.1000-7423.2025.02.006
| [1] | Barnett R, Schistosomiasis[J]. Lancet, 2018, 392(10163): 2431. |
| [2] | Ewuzie A, Wilburn L, Thakrar DB, et al. Association of current Schistosoma mansoni, Schistosoma japonicum, and Schistosoma mekongi infection status and intensity with periportal fibrosis: A systematic review and meta-analysis[J]. Lancet Glob Health, 2025, 13(1): e69-e80. |
| [3] | Grover E, Paull S, Kechris K, et al. Predictors of bovine Schistosoma japonicum infection in rural Sichuan, China[J]. Int J Parasitol, 2022, 52(8): 485-496. |
| [4] | Du CH, Yang H, Yang MX, et al. Assessment of schistosomiasis transmission risk in Nanjian County of Yunnan Province in 2019[J]. Chin J Schisto Control, 2020, 32(5): 531-533. |
| [5] | Shen J, Yu SF, Peng M, et al. iNOS is essential to maintain a protective Th1/Th2 response and the production of cytokines/chemokines against Schistosoma japonicum infection in rats[J]. PLoS Negl Trop Dis, 2022, 16(5): e0010403. |
| [6] | Guo QH, Chen C, Zhou KK, et al. Evaluation of a real-time PCR assay for diagnosis of schistosomiasis japonica in the domestic goat[J]. Parasit Vectors, 2020, 13(1): 535. |
| [7] | Vuong NL, Le Duyen HT, Lam PK, et al. C-reactive protein as a potential biomarker for disease progression in dengue: A multi-country observational study[J]. BMC Med, 2020, 18(1): 35. |
| [8] | Lanza GR, Upatham S, Chen A. A Place-based conceptual model (PBCM) of Neotricula aperta/Schistosoma mekongi habitat before and after dam construction in the Lower Mekong River[J]. PLoS Negl Trop Dis, 2023, 17(10): e0011122. |
| [9] | Liu MM, Feng Y, Yang K. Impact of micro-environmental factors on survival, reproduction and distribution of Oncomelania hupensis snails[J]. Infect Dis Poverty, 2021, 10: 47. |
| [10] | 许晓娟, 陈雪峰, 吴凡, 等. 不同病原学方法检测野鼠日本血吸虫感染效果比较[J]. 中国血吸虫病防治杂志, 2023, 35(6): 573-582, 589. |
| Xu XJ, Chen XF, Wu F, et al. Comparison of the efficiency of different etiological assays for detection of Schistosoma japonicum infections in wild mice[J]. Chin J Schisto Control, 2023, 35(6): 573-582, 589. (in Chinese) | |
| [11] | Tchami Mbagnia MC, Melachio Tanekou TT, Kengne Fokam AC, et al. PCR-based molecular identification of two intermediate snail hosts of Schistosoma mansoni in Cameroon[J]. Parasit Vectors, 2020, 13(1): 158. |
| [12] | Halili S, Grant JR, Pilotte N, et al. Development of a novel real-time polymerase chain reaction assay for the sensitive detection of Schistosoma japonicum in human stool[J]. PLoS Negl Trop Dis, 2021, 15(10): e0009877. |
| [13] | Qin ZQ, Xu J, Feng T, et al. Field evaluation of a loop-mediated isothermal amplification (LAMP) platform for the detection of Schistosoma japonicum infection in Oncomelania hupensis snails[J]. Trop Med Infect Dis, 2018, 3(4): 124. |
| [14] | MacGregor SR, McManus DP, Sivakumaran H, et al. Development of CRISPR/Cas13a-based assays for the diagnosis of schistosomiasis[J]. EBioMedicine, 2023, 94: 104730. |
| [15] | 唐春莲, 申志琴, 雷家慧, 等. CD4+CD25+调节性T细胞对日本血吸虫病疫苗保护性效果的影响及机制研究[J]. 中国血吸虫病防治杂志, 2016, 28(3): 269-274. |
| Tang CL, Shen ZQ, Lei JH, et al. Effect and mechanism of CD4+CD25+ regulatory T cells on protective efficacy of protein vaccine against Schistosoma japonicum in mice[J]. Chin J Schisto Control, 2016, 28(3): 269-274. (in Chinese) | |
| [16] | 李梦茹, 秦志强, 殷堃, 等. 基于环介导等温扩增技术及规则成簇间隔短回文重复序列的日本血吸虫核酸检测方法的建立及评价[J]. 中国热带医学, 2023, 23(7): 686-691. |
| Li MR, Qin ZQ, Yin K, et al. Establishment and evaluation of a LAMP-CRISPR-based nucleic acid detection method for Schistosoma japonicum[J]. China Trop Med, 2023, 23(7): 686-691. (in Chinese) | |
| [17] | 冯婷, 秦志强, 许静, 等. 环介导等温扩增法检测粪样中日本血吸虫虫卵DNA的效果评估[J]. 中国寄生虫学与寄生虫病杂志, 2017, 35(3): 230-234. |
| Feng T, Qin ZQ, Xu J, et al. Efficacy evaluation of a loop mediated isothermal amplification technique in detection of DNA of Schistosoma japonicum eggs in fecal samples[J]. Chin J Parasitol Parasit Dis, 2017, 35(3): 230-234. (in Chinese) | |
| [18] | 周晓农, 朱泽林, 涂宏, 等. 《加快实现消除血吸虫病目标行动方案(2023—2030年)》解读[J]. 中国血吸虫病防治杂志, 2024, 36(1): 7-12. |
| Zhou XN, Zhu ZL, Tu H, et al. Interpretation of the action plan to accelerate the elimination of schistosomiasis in China (2023-2030)[J]. Chin J Schisto Control, 2024, 36(1): 7-12. (in Chinese) | |
| [19] | 吕超, 许晓娟, 杜春红, 等. 《野鼠血吸虫感染情况监测工作方案(2024年版)》解读[J]. 热带病与寄生虫学, 2024, 22(4): 193-197, 216. |
| Lü C, Xu XJ, Du CH, et al. Interpretation of the surveillance plan for Schistosoma japonicum infection in wild rats (2024 edition)[J]. J Trop Dis Parasitol, 2024, 22(4): 193-197, 216. (in Chinese) | |
| [20] | 汪峰峰, 章乐生, 尹晓梅, 等. 安徽省山丘型血吸虫病流行区野鼠血吸虫感染状况调查[J]. 医学动物防制, 2025, 41(1): 1-4, 10. |
| Wang FF, Zhang LS, Yin XM, et al. Investigation on the infection status of schistosomiasis in wild mice in hilly schistosomiasis endemic regions of Anhui Province[J]. J Med Pest Control, 2025, 41(1): 1-4, 10. (in Chinese) | |
| [21] | 梁莎, 曾凡胜, 秦志强. 环境水体中血吸虫尾蚴检测技术的应用研究进展[J]. 寄生虫与医学昆虫学报, 2022, 29(2): 120-125, 136. |
| Liang S, Zeng FS, Qin ZQ. Progress of detection technology of schistosome cercariae from environmental water[J]. Acta Parasitol Med Entomol Sin, 2022, 29(2): 120-125, 136. (in Chinese) | |
| [22] | Phusantisampan T, Rawiwan P, Roy SRK, et al. Reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the specific and rapid detection of Tilapia lake virus[J]. JoVE, 2020(159): e61025. |
/
| 〈 |
|
〉 |