SHORT COMMUNICATIONS

Detection of Trichomonas vaginalis by PCR and LAMP

  • Li-li ZHAO ,
  • Xu MIAO ,
  • Huan-fei XIE ,
  • Bi-han LIU ,
  • Lin YUAN ,
  • Lin LIN ,
  • Shuang HUANG ,
  • Xu-gan JIANG ,
  • Sheng-xia CHEN ,
  • Yu-juan SHEN ,
  • Jian-ping CAO
Expand
  • 1 Department of Laboratory Medicine, School of Medicine, Jiangsu University, Zhenjiang 212013, China
    2 National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention (Chinese Center for Tropical Diseases Research); NHC Key Laboratory of Parasite and Vector Biology (National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention), Shanghai 200025, China

Received date: 2020-08-20

  Revised date: 2020-11-15

  Online published: 2021-04-30

Supported by

National Parasitic Resource Center(TDRC-2019-194-30);National Science and Technology Major Project(2018ZX10713001-004)

Abstract

To establish PCR and LAMP method for detection of Trichomonas vaginalis, the primers were synthesized, and the DNA of cultured of T. vaginalis was amplified. The feasibility of the two methods in clinical application was compared by detecting 40 T. vaginalis-positive and 40 T. vaginalis-negative samples of vaginal discharge identified by clinical microscopy. The specificity of the two methods was analyzed by amplifying DNA of Mycoplasma urealytium, Giardia lamblia, Staphylococcus aureus, Candida albicans and Escherichia coli. The sensitivity of the two methods was analyzed by determining the concentration of cultured T. vaginalis DNA after serial dilution. The results showed that PCR amplification of T. vaginalis DNA resulted in a 263-bp specific band, and the LAMP method caused a change of amplification product color from red to yellow, presenting the DNA ladder on agarose gel electrophoresis. The feasibility results showed that the coincidence rate of the 40 positive and 40 negative clinical samples by PCR were 100% and 100%, while LAMP method resulted in 100% and 97.5%, respectively. The specificity results showed that the DNA of 5 other pathogens could not be amplified by the two methods. The sensitivity test showed that the lower detection limit of the PCR method was 0.5006 ng/μl and the LAMP method 0.05006 ng/μl. These results indicate that the both methods are highly specific and the LAMP is superior to PCR in sensitivity.

Key words: Trichomonas vaginalis; PCR; LAMP

Cite this article

Li-li ZHAO , Xu MIAO , Huan-fei XIE , Bi-han LIU , Lin YUAN , Lin LIN , Shuang HUANG , Xu-gan JIANG , Sheng-xia CHEN , Yu-juan SHEN , Jian-ping CAO . Detection of Trichomonas vaginalis by PCR and LAMP[J]. CHINESE JOURNAL OF PARASITOLOGY AND PARASITIC DISEASES, 2021 , 39(2) : 260 -264 . DOI: 10.12140/j.issn.1000-7423.2021.02.023

References

[1] Aquino M, Hinderfeld AS, Simoes-Barbosa A. Trichomonas vaginalis[J]. Trends Parasitol, 2020,36(7):646-647.
[2] Edwards T, Burke P, Smalley H, et al. Trichomonas vaginalis: clinical relevance, pathogenicity and diagnosis[J]. Crit Rev Microbiol, 2016,42(3):406-417.
[3] Noh CS, Kim SS, Park SY, et al. Comparison of two PCR assays for Trichomonas vaginalis[J]. Korean J Parasitol, 2019,57(1):27-31.
[4] Lan L, Wang PP, Fang DD, et al. Study on polymerase chain reaction for detection of Trichomonas vaginalis[J]. J Changzhi Med Coll, 2015,29(6):409-412. (in Chinese)
[4] ( 兰龙, 王盼盼, 方丹丹, 等. 聚合酶链反应技术检测阴道毛滴虫的研究[J]. 长治医学院学报, 2015,29(6):409-412.)
[5] Rumyantseva T, Golparian D, Nilsson CS, et al. Evaluation of the new AmpliSens multiplex real-time PCR assay for simultaneous detection of Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma genitalium, and Trichomonas vaginalis[J]. APMIS, 2015,123(10):879-886.
[6] Leli C, Castronari R, Levorato L, et al. Molecular sensitivity threshold of wet mount and an immunochromatographic assay evaluated by quantitative real-time PCR for diagnosis of Trichomonas vaginalis infection in a low-risk population of childbearing women[J]. Infez Med, 2016,24(2):112-116.
[7] Ziaei HH, Taghavi M, Fakhar M, et al. Direct diagnosis of Trichomonas vaginalis infection on archived pap smears using nested PCR[J]. Acta Cytol, 2015,59(1):104-108.
[8] Reyes JC, Solon JA, Rivera WL. Development of a loop-mediated isothermal amplification assay for detection of Trichomonas vaginalis[J]. Diagn Microbiol Infect Dis, 2014,79(3):337-341.
[9] Goo YK, Shin WS, Yang HW, et al. Loop-mediated isothermal amplification targeting actin DNA of Trichomonas vaginalis[J]. Korean J Parasitol, 2016,54(3):329-334.
[10] Jiang HQ, Wei SS, Wu LM, et al. Comparison of PCR detection of different primers for Trichomonas vaginalis[J]. J Mod Med Health, 2015,31(22):3448-3450. (in Chinese)
[10] ( 江海强, 魏帅帅, 吴腊梅, 等. 阴道毛滴虫不同引物PCR检测结果比较[J]. 现代医药卫生, 2015,31(22):3448-3450.)
[11] Adao DE, Rivera WL. Loop-mediated isothermal amplification (LAMP) assay for the rapid detection of the sexually-transmitted parasite, Trichomonas vaginalis[J]. Ann Parasitol, 2016,62(1):25-31.
[12] Fichorova R, Fraga J, Rappelli P, et al. Trichomonas vaginalis infection in symbiosis with Trichomonasvirus and Mycoplasma[J]. Res Microbiol, 2017,168(9/10):882-891.
[13] Kuang GR, Yi FY, Yang YJ, et al. Investigation on symbiosis of Mycoplasma hominis in Trichomonas vaginalis in some areas of Guizhou province[J]. J Guiyang Med Coll, 2015,40(3):249-252. (in Chinese)
[13] ( 匡贵榕, 衣凤芸, 杨宇箭, 等. 贵州省部分地区阴道毛滴虫与人型支原体共生情况[J]. 贵阳医学院学报, 2015,40(3):249-252.)
[14] Chen Y, Liu SL, Ma DX, et al. Analysis of 19 342 cases of Trichomonas vaginalis[J]. J Trop Med, 2019,19(9):1177-1179. (in Chinese)
[14] ( 陈杨, 刘素伶, 马丹霞, 等. 19 342例阴道毛滴虫检查结果分析[J]. 热带医学杂志, 2019,19(9):1177-1179.)
[15] Nikpay S, Otaghi M, Azami M, et al. Trichomonas vaginalis infection among women attending laboratory centers in Ilam, Iran[J]. Infect Disord Dug Targets, 2020,20(1):98-101.
[16] Mori Y, Kanda H, Notomi T. Loop-mediated isothermal amplification (LAMP): recent progress in research and development[J]. J Infect Chemother, 2013,19(3):404-411.
[17] Mori Y, Notomi T. Loop-mediated isothermal amplification (LAMP): a rapid, accurate, and cost-effective diagnostic method for infectious diseases[J]. J Infect Chemother, 2009,15(2):62-69.
[18] Yuhua L, Shuai W, Haoran L, et al. Development of a convenient detection method for Trichomonas vaginalis based on loop-mediated isothermal amplification targeting adhesion protein 65[J]. BMC Infect Dis, 2020,20:319-326.
Outlines

/

〈 〉