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. 2023 May 18;14:1165505. doi: 10.3389/fendo.2023.1165505

Table 3.

Overview of the current methodologies for testing small fibres in DSPN [adapted from Sharma S et al. (9)].

Functional tests for SFN
Test Type Technique Equipment Needed Time to acquire results Normative Data Operating Characteristics for DSPN Limitations
Quantitative sensory testing (QST) (41) Non-invasive, quantitative Computerised measurement of thermal thresholds and heat pain thresholds Computerised assessment device, temperature controlled laboratory and a trained technician Takes about half an hour but could take longer, depending on subject concentration Commercial normative data present from the bigger manufacturers. None available Psychophysical test- results are dependent on subject compliance and attention. Complex testing protocols present. Varying reproducibility depending on experience of the unit undertaking testing.
Laser Doppler imager Flare (LDIflare) (42) Non-invasive, quantitative ( Figure 2 ) Measurement of the axon-reflex mediated flare response as a marker of small fibre function Laser Doppler imager, temperature controlled room, operator with experience Image acquisition took ~1 hour with the older method. Newer method takes approximately 25 minutes. Results available immediately One site normative values determined at a single centre. Larger data set of normative valves desired (43) For the newer technique: Sensitivity of 70-75%, specificity of 66-85%, positive predictive value of 74%, and negative predictive value of 86% Dependent on the microcirculation. Patients need to have no significant macrovascular distal circulatory impairment. Has correlation with Confocal microscopy (44).
Current perception threshold (CPT) (45) Non-invasive, quantitative Low current intensity stimulation of the small nerve fibres at frequency of 250 Hz for A-delta fibres and the
5 Hz for C-fibres.
Neurometer device temperature controlled room and a trained technician Takes about half an hour but could take longer, depending on subject concentration None available. Most studies have included age matched controls for comparison. None available Requires active patient co-operation. Like QST, therefore reproducibility has been a challenge and other methodological challenges persist (such as what frequency to use. Not widely available.
Contact Heat Evoked Potentials (CHEPs) (46) Non-invasive, quantitative Measure cerebral responses to thermal stimuli mediated by A-delta fibres Needs thermal threshold testing first. Then small discs are placed on the head to record signals received to the brain from application of 10 to 20 short (a fraction of a second) heat or cold
stimuli at a particular point of interest (face, arm or leg)
Takes about half an hour but could take longer, depending on subject concentration Multicentre normative data on 226 adult subjects are available The AUC for DSPN detection in a small sample has been estimated at 0.778. Requires active patient co-operation. Like QST and CPT, therefore reproducibility has been a challenge. Not widely available. Also unclear if both A-delta and C-fibres are assessed.
Microneuro-graphy (47) Minimally invasive, semi quantitative Measurement of Single fibre recordings from peripheral axons Skilled operator and extensive equipment list. Preserve of a large neurophysiology lab rather than clinic based procedure. May take up to 3 hours to get a satisfactory recording. None available None available. Considered by EFNS to possess grade A evidence for assessing function of the A-delta fibre pathways in patients with neuropathic pain Still primarily a research tool. May have a role in assessment of neuropathic pain rather than early neuropathy. Expensive and needs skill to elicit responses. Patient cooperation is also extensively required.
Laser Evoked Potentials (LEPs) (48) Non-invasive, quantitative Radiant heat generate by laser selectively excites free nerve endings in the superficial skin layers activating nearby A-delta and C -fibre nociceptors CO2-laser stimulator, technician with experience and a temperature controlled room ideally. May take up to 1 hour to complete the procedure and ensure no artefacts presents in readings gained Single centre normative values available on 100 subjects. No decade specific data reported. None available. Studies have used age matched control data. Limited availability. May be useful in demonstrating reduced function but unable to detect enhanced transmission as found in hyperalgesia. Small changes in pain sensitivity are not easily detectable with LEP
Quantitative sudomotor axon reflex test (QSART) (49) Sudomotor Non-invasive, quantitative Information on skin autonomic function and evaluation of postganglionic sudomotor function using acetylcholine iontophoresis Purpose built lab, iontophoresis and sudomotor quantification equipment. 45-60 minutes to complete. Normative data available from specific centres for QSART. A commercially available device QSWEAT is also available No specific data available for DSPN but has been widely used, especially in the Rochester Diabetic Neuropathy study Requires precautions for electrical safety and small risk of minor local injury to the skin
Thermo-regulatory sweat test (TST) (50) Sudomotor
Non-invasive, semi-quantitative
When core temperature
rises beyond a hypothalamic thermoregulatory set point (>38°C),
sweating occurs
Needs a laboratory and a digital camera 90-120 minutes to perform correctly.
Maximal sweating is achieved within 30–65 minutes.
Unclear Helpful data on the TST available in DSPN mainly from the autonomic lab at the Mayo Clinic, Rochester USA. Patients may not be able to tolerate 60 minutes of warming up
Sympathetic Skin Response (51) Sudomotor Non-invasive, quantitative Information on skin autonomic function and evaluation of postganglionic sudomotor function using electrodermal activity Purpose built lab, SSR equipment includes electrodes. 45-60 minutes to complete. Normative data available from specific centres but usually has been derived from a small normative group Minimal data only available in DSPN. Some helpful data in diabetic autonomic neuropathy and bladder dysfunction. Limited availability, needs expertise and experience to test correctly. Popular in Japan.
Sudoscan® (32) Sudomotor Non-invasive, quantitative Testing is based on stimulation of sweat glands by a low-voltage current (<4volts) representing a electrochemical reaction between electrodes and chloride ions, Just the Sudoscan® device Takes less than 5 minutes Comes with inbuilt normative data. Limited experience at the moment Increasing literature now available of its use in DSPN. Similar AUC as IENFD (0.761) in one study. For Cardiac autonomic neuropathy sensitivity was 65%, specificity 80%. Still limited availability. Needs more detailed validation work for different ethnicities.
Neuropad® (52) Simple qualitative indicator of sudomotor dysfunction Simple sticker which changes colour in the presence of sweating. Cheap and easy to avail. Takes less than 10 minutes Qualitative, does not need normative data. Lots of available literature and has been validated against IENFD. In one study, Neuropad had a sensitivity 85% and specificity of 45% for detection of clinical DSPN. Difficult to interpret when there is partial change in colour though. One centre has published data on semi-quantification using digital imaging of the Neuropad®.
Structural tests for SFN
Skin Biopsy (53) Invasive (minimally), quantitative Measurement of intra-epidermal nerve fibre density Sterile equipment for biopsy, access to trained personnel and laboratory Procedure takes 5-10 minutes but takes a few days to get the results back. Worldwide normative Data present Published sensitivity doe DSPN is between 60% and 95% and specificity between 90% and 95% Challenging to use in prospective studies of very large cohorts, infection risk at site of biopsy
Sural nerve biopsy (54) Invasive, quantitative Ultrastructure and morphometric analysis of sural nerve biopsy specimens Experienced operator who can perform biopsy and access to pathologist and at times, electron or confocal microscope Procedure may take up to 45 minutes. Results usually take a few days. None available None available Infection, pain and hypoesthesia at biopsy site
Corneal Confocal Microscopy (55, 56) Non-invasive, quantitative Measurement of nerve parameters of the corneal sub-basal layer Corneal scanning confocal microscope, trained technician Image acquisition takes 5-10 minutes. Results available immediately if automated counting used Worldwide normative Data present Reported sensitivity
of 85% and specificity of 84%
Surrogate marker of DSPN rather than a direct indicator. Previously reliant on manual counting but newer automated methods emerging. Unclear which of the three- CNFB, CNFL or CBNFD best representative/predictive of DSPN