Utility of Multi-target Nested PCR and ELISPOT Assays for the Detection of Paucibacillary Leprosy: A Possible Conclusion of Clinical Laboratory Misdiagnosis

Frontiers in Cellular and Infection Microbiology

Zhenzhen Wang¹  Limei Wu¹  Hongsheng Wang³  Siyu Long⁶  Qiang Yao⁶  Santosh Chokkakula⁶  Heng Long⁶  Ying Shi⁶  Wenming Kong⁷  Lihua Hu⁷  Haiqin Jiang⁷  Wenyue Zhang⁷ 

[1] Peking Union Medical College, Nanjing, China;Centre for Global Health, School of Public Health, Nanjing Medical University, Nanjing, China;Department of Leprosy Control, Wenshan institute of Dermatology, Wenshan, China;Department of Leprosy Control, Zhejiang, Provincial Institute of Dermatology, Zhejiang, China;Department of Microbiology, Chungbuk National University College of Medicine, and Medical Research Institute, Cheongju, South Korea;;Department of Mycobacterium, Jiangsu Key Laboratory of Molecular Biology for Skin Diseases and STIs, Institute of Dermatology, Chinese Academy of Medical Sciences &National Centre for STD and Leprosy Control, China CDC, Nanjing, China;

关键词: diagnosis;    PCR;    nested PCR;    ELISPOT;    paucibacillary leprosy;   

DOI  :  10.3389/fcimb.2022.814413

来源: DOAJ

【 摘 要 】

The diagnosis of paucibacillary (PB) leprosy often possesses a diagnostic challenge, especially for pure neuritic and lesser skin lesions with the zero bacillary load, requiring a sensitive and accurate diagnostic tool. We have included 300 clinically diagnosed new leprosy cases (comprising 98 PB cases) and analyzed the sensitivity and specificity of PB leprosy cases by nested PCR with folP, gyrA, rpoB, RLEP, and 16SrRNA and Enzyme-linked Immunospot Assay test (ELISPOT) with MMPII, NDO-BSA, and LID-1 antigens by detecting interferon gamma (IFN-γ) release. The overall positivity rates of genes tested in 300 clinical specimens were identified as 55% of 16SrRNA, 59% of RLEP, 59.3% of folP, 57.3% of rpoB, 61% of gyrA while 90% of nested folP, 92.6% of nested rpoB, and 95% of nested gyrA, and 285 (95%) of at least one gene positive cases. For PB specimens, 95% PCR positivity was achieved by three tested genes in nested PCR. The data obtained from ELISPOT for three antigens were analyzed for IFN-γ expression with 600 subjects. Among 98 PB leprosy cases, the sensitivity of MMP II, LID-1, and NDO-BSA was 90%, 87%, and 83%, respectively, and the specificity was 90%, 91%, and 86%, respectively. The total number of cases positive for at least one antigen was 90 (91.8%) in PB, which is significantly higher than that in multibacillary (MB) leprosy (56.7%). The combination of multi-targets nested PCR and ELISPOT assay provides a specific tool to early clinical laboratory diagnosis of PB leprosy cases. The two assays are complementary to each other and beneficial for screening PB patients.

【 授权许可】

Unknown