Real-time loop-mediated isothermal amplification (real-time LAMP) assay for rapid diagnosis of amoebic liver abscess

被引:3
作者
Khunger, Sandhya [1 ,2 ]
Mewara, Abhishek [1 ]
Kaur, Upninder [1 ]
Duseja, Ajay [3 ]
Ray, Pallab [4 ]
Kalra, Naveen [5 ]
Sharma, Navneet [6 ]
Sehgal, Rakesh [7 ]
机构
[1] Postgrad Inst Med Educ & Res, Dept Med Parasitol, Chandigarh, India
[2] Shree Guru Gobind Singh Tricentanary Univ, Dept Microbiol FAHS, Gurugram, India
[3] Postgrad Inst Med Educ & Res, Dept Hepatol, Chandigarh, India
[4] Postgrad Inst Med Educ & Res, Dept Med Microbiol, Chandigarh, India
[5] Postgrad Inst Med Educ & Res, Dept Radiodiag & Imaging, Chandigarh, India
[6] Postgrad Inst Med Educ & Res, Dept Internal Med, Chandigarh, India
[7] Aarupadai Veedu Med Coll & Hosp, Vinayaka Missions Res Fdn DU, Pondicherry, India
关键词
amoebic liver abscess (ALA); conventional PCR; Entamoeba histolytica; real-time LAMP; real-time PCR; ENTAMOEBA-HISTOLYTICA DNA; PCR; DISPAR;
D O I
10.1111/tmi.13954
中图分类号
R1 [预防医学、卫生学];
学科分类号
1004 ; 120402 ;
摘要
Among the parasitic diseases, amoebic liver abscess (ALA) ranks second to malaria in terms of mortality. Due to the poor sensitivity of conventional diagnostic methods, there is a need for the development of effective and rapid diagnostic methods for ALA. Thus, the purpose of this work was to develop a real-time loop-mediated isothermal amplification (RT-LAMP) assay specific to Entamoeba histolytica. Further, we compared the performance of real-time LAMP with conventional and real-time PCR (RT-PCR) targeting 18S small subunit ribosomal RNA (18S SSU rRNA) gene of E. histolytica in patients with ALA. A total of 126 liver samples were obtained for the study. Of these, 96 aspirated pus samples were obtained from patients suffering from an ALA (serology confirmed, anti-amoebic immunoglobulin IgG positive), 19 aspirated pus samples from patients with pyogenic liver abscess (PLA, 16S RNA gene positive) and 11 autopsy liver tissues. The results showed that the DNA of E. histolytica was detected in 81 samples by conventional PCR, 93 by RT-PCR and 95 by RT-LAMP. The analytical sensitivity of the RT-LAMP assay was much higher than the other two techniques. RT-LAMP assay was able to amplify up to one copy of the targeted gene of E. histolytica while conventional PCR and RT-PCR could amplify up to 10(3) and 10(2) copies of the targeted gene of E. histolytica, respectively. In conclusion, RT-LAMP proved to be a sensitive, specific and rapid test which can be utilised as an effective tool for the diagnosis of ALA.
引用
收藏
页码:104 / 112
页数:9
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