Use of fine needle aspirate from peripheral nerves of pure-neural leprosy for cytology and PCR to confirm the diagnosis: A pilot study

Reja, Abu Hena Hasanoor; De, Abhishek; Biswas, Supratik; Chattopadhyay, Amitabha; Chatterjee, Gobinda; Bhattacharya, Basudev; Sarda, Aarti; Aggarwal, Ishad

doi:10.4103/0378-6323.120731

Cited by 13 publications

(6 citation statements)

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“…The basic premise of the thought here was that FNA is a much simpler technique, which does not require a high level of skill and can be done routinely in OPD set up as a day care procedure. [ 3 ] However, the sample size in this study was only 13 patients. To check the reliability, reproducibility and precision of FNA combined with ZN staining and multiplex-PCR in the diagnosis of PNL, the authors performed these tests on a larger sample size of 47 patients over a span of 3 years in Dermatology OPD of a tertiary care center in eastern India.…”

Section: Discussionmentioning

confidence: 99%

“…However, nerve biopsy being the major deterrent in our efforts to diagnose PNL, we had conducted a pilot study where we demonstrated M. Leprae DNA in specimens obtained out of FNA done on affected peripheral nerves in 13 patients. [ 3 4 ] FNA, being a relatively less invasive and simpler procedure, could be done with ease in our outpatient set up. The diagnostic yield of FNAC-PCR was as high as 84.6% in the pilot study.…”

Section: Discussionmentioning

confidence: 99%

“…In an effort to find a simple yet objective and test to diagnose PNL, the authors demonstrated in a pilot study that fine needle aspiration cytology (FNAC) when combined with Ziehl-Neelsen (ZN) staining and PCR gives a good diagnostic yield. [ 3 4 ] FNAC being a less invasive test needs less skill and infrastructure and comes with lesser rates of complications and can be easily performed within the existing infrastructure. With this background and in pursuit of further evidence, the authors conducted this study over a larger sample size, for a longer duration to determine the reliability, efficacy, and reproducibility of FNAC combined with ZN staining and PCR in diagnosis of PNL.…”

Section: Introductionmentioning

confidence: 99%

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Use of fine needle aspirate from peripheral nerves of pure-neural leprosy for cytology and polymerase chain reaction to confirm the diagnosis: A follow-up study of 4 years

Reja

Aggarwal

et al. 2017

Indian J Dermatol

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Background:Pure neural leprosy (PNL) still remains a diagnostic challenge because of the absence of sine qua non skin lesions of leprosy and a confirmatory diagnostic method. The authors had earlier described a simple yet objective technique of combining fine needle aspiration cytology (FNAC) coupled with a multiplex polymerase chain reaction (PCR) in a pilot study, wherein the technique showed promise of a reliable diagnostic tool. In the pursuit of further evidence, the authors carried out a 4-year study with PNL cases to find the efficacy and reliability of the said method in a larger sample size.Aim:This study was conducted to find the efficacy, reliability, and reproducibility of FNAC coupled with multiplex PCR and Ziehl-Neelsen (ZN) staining in identifying the cases of PNL.Materials and Methods:All cases that were suspected to be suffering from PNL, following evaluation by two independent observers were included in the study and were subjected to FNAC from the affected nerve, and the aspirates were evaluated for cytology, ZN staining, and multiplex PCR for Mycobacterium leprae genome. In addition, serum anti-PGL1 levels were also performed in all the study subjects. Fifteen non-PNL cases were also included in the control arm.Results:A total of 47 cases were included in the test arm and subjected to FNAC. Conventional ZN staining could demonstrate acid-fast bacilli (AFB) in only 15 out of 47 cases (31.91%) while M. leprae DNA could be elicited in 37 (78.72%) cases by the multiplex PCR. Only 13 (27.65%) out of 47 cases showed anti-PGLI-1 antibody positivity. On cytological examination of the nerve aspirates, only 11 (23.40%) cases showed epithelioid cells whereas nonspecific inflammation was seen in 26 (75.60%) cases.Conclusion:The results of this study conducted over a larger sample size corroborate with the findings of our pilot study. In a resource poor set up, FNAC in combination with ZN staining and multiplex PCR is a rapid, simple, and easily performed test, which can give a reproducible and objective diagnosis in cases of PNL.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Use of fine needle aspirate from peripheral nerves of pure-neural leprosy for cytology and polymerase chain reaction to confirm the diagnosis: A follow-up study of 4 years

Reja

Aggarwal

et al. 2017

Indian J Dermatol

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“…Recently, the authors have described a simple but objective method of diagnosis of PNL by using fine needle aspiration and multiplex PCR. [ 4 ] However, we perceive that there is a considerable lack of data regarding the genomic changes in M. leprae which confines itself to a low grade infection of peripheral nerves for years. We postulate that this adaptation to this intra-cellular niche may be accompanied by either gene loss or mutation in susceptibility genes.…”

Section: Introductionmentioning

confidence: 99%

Unique TTC repeat base pair loss mutation in cases of pure neural leprosy: A survival strategy of Mycobacterium leprae?

Reja

Biswas

et al. 2015

Indian J Dermatol

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Background:Genomic reduction helps obligate intracellular microbes to survive difficult host niches. Adaptation of Mycobacterium leprae in cases of pure neural leprosy (PNL) in the intracellular niche of peripheral nerves can be associated with some gene loss. Recently, a stable but variable number of tandem repefzats (TTC) have been reported in strains of M. leprae. FolP and rpoB genes are the two common mutation sites which deal with the susceptibility of the bacteria to drugs.Aim:We attempted to find if genomic reduction of M. leprae in context of these TTC repeats or mutations in folP1 and rpoB can be the reason for the restriction of M. leprae in the nerves in PNL.Materials and Methods:DNA extracts taken from fine needle aspiration of affected nerves of 24 PNL cases were studied for tandem repeats with 21TTC primer in multiplex-PCR. Mutations were also studied by PCR Amplification of SRDR (Sulphone Resistance Determining Region) of the folP1 and multiple primer PCR amplification refractory mutation system (MARS) of the rpoB.Results:Of the 24 PNL, only 1 patient showed mutation in the rpoB gene and none in the folp1 gene. Studying the mutation in TTC region of the M. leprae gene we found that all the cases have a loss of a few bases in the sequence.Conclusion:We can conclude that there is consistent loss in the bases in the TTC region in all cases of pure neural Hansen and we postulate that it may be an adaptive response of the bacteria to survive host niche resulting in its restriction to peripheral nerves.

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“…[ 5 6 7 ] A comprehensive view of the issues involved has not been detailed in any of them or in other publications on this subject. [ 8 9 ] This retrospective study was undertaken to analyze the feasibility, efficacy, and role of FNA cytology of sensory cutaneous nerves in Hansen's disease.…”

Section: Introductionmentioning

confidence: 99%

Sensory cutaneous nerve fine-needle aspiration in Hansen′s disease: A retrospective analysis of our experience

Prasoon

Mandal

Agrawal

2015

J Cytol

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Background:Leprosy affects peripheral nerves. As Mycobacterium leprae has unique tropism for Schwann cells, thickened sensory cutaneous nerves provide an easy target for the detection of lepra bacilli and other changes associated with the disease.Materials and Methods:The data of patients with sensory cutaneous nerve involvement were retrieved from our record for the period January 2006 to December 2014. The hematoxylin and eosin (H and E)- and May-Grünwald-Giemsa (MGG)-stained slides were screened for Schwann cells, granuloma, and necrosis. Modified Ziehl-Neelsen (ZN)-stained smears were searched for lepra bacilli and globi. Morphological index was calculated in multibacillary lesions.Result:Twenty-nine sensory cutaneous nerves were aspirated in 23 patients. While 15 cases showed skin and nerve involvement, 8 cases showed only nerve involvement. Terminal cutaneous branch of the radial nerve was most often aspirated. No motor loss was observed after aspiration. Five cytologic pictures were seen — Epithelioid cell granuloma only in 6 cases, epithelioid cell granuloma with necrosis in 1 case, epithelioid cell granuloma with lepra bacilli in 3 cases, necrosis with lepra bacilli in 1 case, and only lepra bacilli in 12 cases. Morphological index ranged from 20% to 80%.Conclusion:Sensory cutaneous nerve fine-needle aspiration (FNA) is a feasible, viable, effective, and safe procedure. It adds to diagnostic FNA yield in patients with concomitant skin involvement and offers a way to evaluate patients with only nerve involvement. Calculation of morphological index allows prognostication and may have a role in assessing response to therapy and/or relapse.

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Use of fine needle aspirate from peripheral nerves of pure-neural leprosy for cytology and PCR to confirm the diagnosis: A pilot study

Cited by 13 publications

References 7 publications

Use of fine needle aspirate from peripheral nerves of pure-neural leprosy for cytology and polymerase chain reaction to confirm the diagnosis: A follow-up study of 4 years

Use of fine needle aspirate from peripheral nerves of pure-neural leprosy for cytology and polymerase chain reaction to confirm the diagnosis: A follow-up study of 4 years

Unique TTC repeat base pair loss mutation in cases of pure neural leprosy: A survival strategy of Mycobacterium leprae?

Sensory cutaneous nerve fine-needle aspiration in Hansen′s disease: A retrospective analysis of our experience

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