Dataset Information

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

ABSTRACT: 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.

SUBMITTER: Jiang H

PROVIDER: S-EPMC9036522 | biostudies-literature | 2022

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

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

Jiang Haiqin H Shi Ying Y Chokkakula Santosh S Zhang Wenyue W Long Siyu S Wang Zhenzhen Z Kong Wenming W Long Heng H Wu Limei L Hu Lihua L Yao Qiang Q Wang Hongsheng H

Frontiers in cellular and infection microbiology 20220411

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 ...[more]

PMID: 35480232

Similar Datasets

Project description:BackgroundThe Comprehensive T Cell Vaccine Immune Monitoring Consortium (CTC-VIMC) was created to provide standardized immunogenicity monitoring services for HIV vaccine trials. The ex vivo interferon-gamma (IFN-γ) ELISpot is used extensively as a primary immunogenicity assay to assess T cell-based vaccine candidates in trials for infectious diseases and cancer. Two independent, GCLP-accredited central laboratories of CTC-VIMC routinely use their own standard operating procedures (SOPs) for ELISpot within two major networks of HIV vaccine trials. Studies are imperatively needed to assess the comparability of ELISpot measurements across laboratories to benefit optimal advancement of vaccine candidates.MethodsWe describe an equivalence study of the two independently qualified IFN-g ELISpot SOPs. The study design, data collection and subsequent analysis were managed by independent statisticians to avoid subjectivity. The equivalence of both response rates and positivity calls to a given stimulus was assessed based on pre-specified acceptance criteria derived from a separate pilot study.FindingsDetection of positive responses was found to be equivalent between both laboratories. The 95% C.I. on the difference in response rates, for CMV (-1.5%, 1.5%) and CEF (-0.4%, 7.8%) responses, were both contained in the pre-specified equivalence margin of interval [-15%, 15%]. The lower bound of the 95% C.I. on the proportion of concordant positivity calls for CMV (97.2%) and CEF (89.5%) were both greater than the pre-specified margin of 70%. A third CTC-VIMC central laboratory already using one of the two SOPs also showed comparability when tested in a smaller sub-study.InterpretationThe described study procedure provides a prototypical example for the comparison of bioanalytical methods in HIV vaccine and other disease fields. This study also provides valuable and unprecedented information for future vaccine candidate evaluations on the comparison and pooling of ELISpot results generated by the CTC-VIMC central core laboratories.

Project description:IntroductionThe present study aimed to evaluate leprosy cure and relapse rates as primary outcomes related to two additional strategies for leprosy treatment: clofazimine for paucibacillary (PB) leprosy patients and clarithromycin for patients with rifampicin-resistant leprosy.MethodsWe conducted two systematic reviews (protocols CRD42022308272 and CRD42022308260). We searched the PubMed, EMBASE, Web of Science, Scopus, LILACS, Virtual Health Library and Cochrane Library databases, registers of clinical trial databases and gray literature. We included clinical trials evaluating the addition of clofazimine to PB leprosy treatment and the use of clarithromycin for treating patients with rifampicin-resistant leprosy. Risk of bias (RoB) in randomized clinical trials was assessed by the RoB 2 tool and that in non-randomized clinical trials was assessed by the ROBINS-I tool; and the certainty of the evidence was assessed by the Grading of Recommendations Assessment, Development and Evaluation (GRADE) system. A meta-analysis of dichotomous outcomes was performed.ResultsFor clofazimine, four studies were included. Cure and relapse rates were not different with the addition of clofazimine to PB leprosy treatment and demonstrated very low certainty of evidence. For clarithromycin, six studies were included. Considerable heterogeneity resulted from the difference between comparators, and studies showed no difference in the assessed outcomes with the addition of clarithromycin to rifampicin-resistant leprosy treatment. Mild adverse events were reported for both drugs but did not significantly impact treatment.DiscussionThe effectiveness of both drugs still needs to be determined. Adding clofazimine to PB leprosy treatment may reduce the repercussions of an incorrect operational classification with no apparent relevant side effects.Systematic review registrationhttps://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42022308272; https://www.crd.york.ac.uk/prospero/display_record.php?ID=CRD42022308260, identifier: CRD42022308272; CRD42022308260.

Project description:IntroductionLeprosy is a chronic infectious disease caused by the agent Mycobacterium leprae, characterized by its high disabling power. Data points to Brazil being the second country with the highest number of cases in the world, behind only India, representing a major challenge for public health. This work aims to determine the prevalence of paucibacillary (PB) cases in relation to leprosy cases in Brazil, using data published in the literature.MethodsThis is a systematic review and meta-analysis carried out with studies from the last 20 years, being developed based on the Preferred Reporting Items for Systematic Review and Meta-analyzes (PRISMA).The search was carried out in the databases: PUBMED, SciELO, LILACS (via VHL)and Science Direct in October 2023, using the following descriptors ((("Brazil" [Mesh]) AND ("Leprosy, paucibacillary" [Mesh])) AND "Epidemiology" [Mesh]), in English, Portuguese and Spanish. Original studies of the analytical case-control, cohort, cross-sectional, epidemiological types were selected, as well as articles with satisfactory information for numerical extraction with separate data on the paucibacillary and multibacillary clinical forms. The methodological quality assessment followed the JBI critical appraisal checklist. Statistical analysis was performed using the Comprehensive Meta-Analyses-CMA software, version 3.0 (Biostat, Engewood, NJ, United States).ResultsThe meta-analysis of the 48 studies obtained a paucibacillary prevalence rate in Brazil of 50.5% or 0.505 (95% CI = 0.502-0.509).The differences in the analyzes were statistically significant (Q-value 4302.681;df 47; I 98.905), with a high heterogeneity value evidenced by I2 (98.905). This analysis demonstrated that the frequency in the Midwest region was the highest and the South region was the lowest (21.4%). Begg's (Kendall Tau p = 0.35) and Egger's tests (p = 0.20) were performed, in which no high publication bias was noted. Subgroup analysis indicated that paucibacillary cases varied from region to region, with the Midwest region having the highest prevalence and the South region having the lowest.ConclusionThe results stand out significantly for the research gaps that investigate PB cases, requiring more research aimed at investigating the paucibacillary clinical form that can contribute to the early diagnosis of leprosy.Systematic review registrationPROSPERO code: CRD42024514106.

Dataset Information

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

Publications

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

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets