This document discusses various methods for quantitatively assessing peripheral neuropathy in patients with diabetes. It describes testing vibration perception thresholds using a vibratory instrument, as well as testing heat and cold perception thresholds using a device that applies controlled temperatures. Electrodiagnostic testing such as electromyography and nerve conduction studies are also mentioned as valuable but not always available or affordable methods. The importance of standardized, quantitative testing is discussed for accurately determining neuropathy severity and a patient's risk level.

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Chapter VII
LABORATORY ASSESSMENT
 UNDERSTANDING THRESHOLDS AND DEFICITS
 NEED FOR THEIR QUANTIFICATION
 DESIRABLE QUALITIES IN ASSESSMENT MODALITIES
 TESTING BY MONOFILAMENT
 TESTING FOR AND INTERPRETATION OF VIBRATION THRESHOLDS
 TESTING FOR HEAT AND COLD PERCEPTION THRESHOLDS
 CASE IV SYSTEM, EMG AND NCV
Understanding thresholds and deficits while testing for neuropathy:
Clinical testing of peripheral nervous system gives only a qualitative or at best semi
quantitative assessment. It does not exactly quantify deficits. The greater the deficits
on testing, the more vulnerable the foot becomes to the development of ulcers, which
can get infected and is under a greater chance of getting amputated. Qualitative
testing cannot correlate the findings of clinical testing precisely to identify the level
of risk a foot has. And if this cannot be done, anticipatory measures to prevent the
foot from getting ulcerated would be difficult to initiate.
Need for quantification:
Thus some form of quantitative assessment of the diabetic foot is essential that can
identify the foot at high risk as also the quantum of the risk associated with it as
compared to normal foot. This can be done by semi quantitative methods using
instruments in a graded manner, progressing to more precise quantitative methods.
All the sensations of lower limbs however are not quantifiable. The touch and pain
are qualitative and / or subjective and quantifying them is not without inherent
errors, especially pain. On the other hand a combination of touch-pressure, vibration
and thermal stimuli are highly quantifiable. These can be tested in quantitative
manner by using increasingly precise instruments.
Precautions and care exercised during testing:
There could be inherent errors that may get into the testing, eg to the instruments,
the users, the imperfections on the part of the patients to understand while reporting
and when recorded.
Any test for sensory neuropathy (and for that matter, motor and autonomic
neuropathy) becomes meaningful when attention is paid to:
1. The choice of the evaluation procedure which carries with it, its limitations on
quantification, reproducibility, sensitivity and specificity as compared, with
other tests that bring out similar data.

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2. The tests inherently bring in, the operator or hands on variables and the varying
patient responses that need to be cross checked and controlled.
3. The analysis of the tests performed. The test procedure could still have a subjective
element that can seriously interfere with the accuracy of the data obtained (and
recorded.)
4. The standardization is an important consideration especially if a procedure is
inherently multistage or complicated and depends on subjective responses as in
sensory neuropathy testing. (In motor neuropathy by some contrast the subjective
element is less, as evidenced in electromyography and clinical testing correlations.)
The issue of the standardized outputs (as in say vibration threshold measuring
instruments) or sensitivity of instruments is also an important consideration.
5. The reference values that we may use should be standard, they should be reasonably
simple to understand and usable, even if there normative ranges are multivariate
dependant (like age, duration of diabetes) the distinction of what is normal and what
is not should be clear.
As much as these considerations are valid and valuable in controlled clinical trials or
epidemiological studies, they are important for clinical practice as well. In fact, the
greater the accuracy or sensitivity or reliability of them, the yield of information is as
valuable even if the number of patients that we may analyze are smaller.
Qualities desirable in a modality of testing:
They are the relevance, reliability, specificity and sensitivity of these methods for the
prediction of a foot at high risk.
The laboratory tests for sensory neuropathy: The monofilament, vibration
and heat and cold perceptions:
The “Semmes Weinstein Nylon Monofilament” carries out the test for the combined
sensation of light touch and pressure. The filament is held as indicated on the
filament perpendicular to the sole and is pressed and removed immediately evenly
from the heads of the 1st 3rd and the 5th metatarsals, mid foot and the dorsum of
each foot. The findings are recorded as sensed or not sensed. The test correlates
satisfactorily well with more precise vibration perception thresholds to indicate a
foot is at increased risk. The monofilament most commonly used is the 10 grams
monofilament meaning thereby that it exerts a pressure of 10 grams when pressed
perpendicular to the sole. This quick test can by and large help decide a foot at risk
for further evaluation.
The tests for the sensation of vibration are carried out by vibration threshold
measuring instruments like Sensitometer. The thermal threshold is tested by an
electrical instrument that can apply progressively lowered or elevated measurable
temperatures to the skin, to see the threshold at which, the sensation of pain is
evoked. The usual clinical testing is relatively simple and could be applied easily.
The quantitative testing for detection for hot and cool thresholds would require a
sophisticated instrument. These instruments are not easily available in our country
and DFSI and NNEF have helped successfully develop them and are now
manufactured in India. Following are some details of these two modalities.

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Testing vibration perception:
This is an electrically operated small instrument contained in a box that has now
been developed in India and has been validated against English Instruments as
equally good. It weighs 6 kg (Fig 62). It works on the principle of application of
electrical energy to a calibrated vibrator. This voltage can be increased from 0 to 50 V
by turning the control switch to the right in a clockwise manner gradually. The volts
appear on the top of the semicircle and the vibratory amplitude calibrated as microns
of motion appear below the semicircle. (Fig. 27)
The silent switch is used to detect if the patient is 1) is either not able to understand
the test and what is it that he needs to report, 2) or is trying to accommodate to the
demand of the operator to report if he feels the vibration or not or 3) he is simply
bluffing. It is used to stop the vibrations while testing but does not lower the
indicator from the voltage at which the vibrations were delivered at that time.
The procedure of testing should be explained to the patient in detail to remove any
apprehension and to obtain proper response.
The trial run:
It is preferable to place the vibrator on the fingertips of the hand of the patient to
show the patient the kind of sensation that he is going to get by increasing the
voltage slowly. The feet may then be tested by slowly rotating the switch to right till
the patient gets the sensation and says yes. Increase it a little to see if the sensation
is more pronounced. Then slowly turn back to see whether the patient indicates
absence of sensation little below the point at which he / she had indicated the
sensation for the first time. Increase the voltage again very gradually to see the
voltage at which he again registers the sensation of vibration. Record the voltage for
that point. Then record the voltages at the heads of the first, third and the fifth
metatarsal, the mid foot and the heel of both the feet. All the points tested by the S W
Monofilament should also be tested for the vibration perception threshold.
Record voltage at each point in the data record form. In one of the instrument
available the multiple voltages recorded can be shown in a graphical for like an
audiometric measurement.
It is important to tell the patients to report the earliest and the most minimal of the
sensation of vibration, as soon they are sure. Otherwise, they tend to wait till it is
quite strong. That will give an erroneously high reading.
The single reading to be entered is the mean of two or three readings done at that
point and should be nearly the same or less than 10% away from each other.
The factors that may affect the threshold of vibration:
Age: With increasing age the threshold goes on increasing, above the age of 50.
The lower extremities: The nerves of the lower extremities appear to be more
vulnerable to increasing threshold as age advances. The longer the length of the
nerve fibre the more it may be susceptible to loss of vibration giving rise to higher

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threshold. In person with diabetes, the threshold for vibration is more likely to be
higher than the normal individuals even in the upper extremities.
Fatigue: Fatigue usually raises the vibration perception threshold,
Muscle tone: An inverse relationship is found to exist between the muscle tone and
the Vibration Perception Thresholds. Therefore, it is recommended that the testing is
carried out in a single position with patient resting and relaxing for 5 to 10 minutes
before testing.
Neuropathy: All such factors that cause neuropathy e.g diabetes mellitus, will cause
an elevation in VPT.
Some important interrelations of thresholds and neuropathic abnormalities:
1. The vibration perception thresholds (VPTs) are significantly increased in both upper
and lower limbs in persons with diabetes patient.
2. The VPTs for great toe is higher and higher VPTs of great toes are associated
positively with longer duration of diabetes.
3. VPTs values positively correlate with observed sensory loss and areflexia, correlate
with increasingly severe stages of person with diabetes neuropathy.
4. One abnormality of quantitative sensory testing correlates well with some other form
of quantitative testing. For example vibration and cooling thresholds were strongly
associated with clinical deficits.
5. The vibration and cooling threshold help establish minimum criteria to diagnose
polyneuropathy.
A sample of report that can be given to the patient is attached above in the section on
the clinical and laboratory assessment for vibration perception threshold.
Interpreting the values of the VPT:
It is not the first value on a patient that is the most important observation. It is
the direction in which the second value appears is important and the
quantum change. We may obtain a second value in the next one year. If the
second or the third values are progressively higher then it is clear that there is
a progressive deterioration in the neuropathic status of the individual. If the
values do not vary much and cluster around the first value, the position
regarding the status of neuropathy is more or less stable. If the change is
unidirectional (or monotone) then higher the value or the quantum of the
change, the greater will it indicates the deterioration to be.
On foot, if a patient fails to detect the vibrations when the voltage applied
exceeds 25V, it is indicative of threefold to fourfold increased risk of
developing foot ulceration. When the threshold for detecting vibration
exceeds 42V the risk rises to nearly 17 fold.

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Testing for sensations of Heat, Cold, Heat and Cold by HCP Sensitometer.
India is a hot country with barefoot walking a common practice. A person with
Diabetic Foot may not sense temperatures as high as engine casing which may result
in to burn injuries. Thus it is logical to verify the integrity of the thermal sensations
like cool warm and pain associated with it, for safety reasons.
Normal ranges of these thresholds:
32 to 340C is neutral zone, just below 320C & up to 300C is the Cool threshold range -
age dependant. The sensation of cool pain appears between 27 to 250C. 36 to 380C is
the interval for the sensation of warmth, 42 to 450C is the heat pain zone. Above
ranges are true for clinical practice in an ambient temperature of 27 to 370C and skin
temperature doesn't affect significantly in clinical practice.
Sensitometer-HCP (Figure 61) is used by many in India since 2003. It offers reliable
and reproducible thresholds. It needs about 15 to 20 minutes of a technician. It
indicates simultaneous possibility of Autonomic Dysfunction. The normal heat pain
threshold range is 42 to 450C, erythema takes place after an hour at 450C, needs 10
seconds to a minute at 500C for tissue damage, and just one second at 550C. When the
heat and cold perceptions are lost in feet, as compared to the hands the patient can
perceive the problem especially when the quantification of the deficit is shown.
A rate of one degree per second is generally recommended by some investigators.
But for heat and cold thresholds the rate of rise or fall of temperature seems best at 1
degree Centigrade over 4 seconds since C fibers carry sensations at a much slower
speed, have a period of latency of about 500 milliseconds before the impulse gets
initiated. Consequently there is a delay in reaching the sensation to the cortical areas.
There is a further delay for the registration of the sensation and response to it. These
factors determine the rate of rise or fall of temperature that the patient needs to sense
and respond to. This makes testing a little longer but gives precise thresholds,
reduces the need for many readings, averaging etc.
Concept of heat pain and cold pain should be understood; these thresholds are
higher and lower than heat and cold respectively.
Cool pain is sensation of coolness with an additional component of un-comfort. Some
thing similar to standing near the AC.
Heat pain is warmth plus pricking sensation.
It is the slight un-comfort to be detected for the thresholds and not the ability to bear
chill or heat.
Case IV System for Neuropathy:
Following is a small simplified review of the research that has gone in, in the
quantitative and clinical methods of investigation in peripheral neuropathy to
answer the issues. Elaborate systems to identify the neuropathic symptom score, the
neuropathy deficit identification or to estimate on analog scales positive symptoms
like pain or hyperesthesia were first worked out. Using computer analyzable codes

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the data was entered and the information was obtained using trained observers in an
unbiased manner. The data consisted of examination findings, as also the
instrumental testing that would give more quantified results. All this was fed to a
“Computer Assisted Sensory Evaluation System IV” and then analyzed for results.
PK Thomas and Peter Dyke were the investigators who developed this system and
used it over long years with continuing improvements. This system tests all the
sensations and the positive symptoms as also the deficits. Dyke and Thomas found
that if any one sensation tested by this system was found to be abnormal then there
would be at least one more sensation that is likely to be abnormal indicating that the
foot is at a higher risk. On further testing the possibility of whether an abnormality
detected by one test could be correlated to the abnormality detected by another test
within the system, it was found that the correlation was good. The question of
quantifying the associated risk in the presence of such findings was answered by
conducting trial in the Rochester population.
Electromyography and nerve conduction velocity:
It is a modality that has become available more routinely in only last few years with
the subspecialty of neurology. For primary physicians and Diabetologists, its routine
use is still expensive, not easily available. That makes the repeat testing for assessing
the efficacy of interventional strategies not only costly but also difficult.
No doubt electromyography and nerve conduction velocity is the gold standard for
assessing neuropathic abnormalities and differentiating them primarily for the
pathologic abnormality of nerve fibers viz. axonal or myelin abnormalities. No other
method of testing neuropathy gives this particular differentiation in such clear terms.
Its use for monitoring is the gold standard as the subjective elements are minimal
and it is only operator controlled. Additionally, it has great value in differentiating
between the purely nerve fiber pathology, vis-à-vis, the possible disconnections of
reflex arc at the spinal level. And lastly its usefulness for assessment of the posterior
columns of spinal cord is valuable to those who deal with spinal cord and vertebral
column problems, though not always for the assessments for Diabetologists.
If there is detectable abnormality in say vibration perception, there will be some
abnormality in the EMG / NCV testing also. Thus suffice it to say that the simpler
tools like vibration threshold measuring instruments we have sought to elaborate
upon in this volume, to be placed in the hands of physician and Diabetologists to
prevent foot problem are like surrogate markers for the EMG / NCV abnormality.
Often therapeutic intervention will cause some of the parameters of EMG and NCV
to improve but that does not always mean a reversal of neural pathologies as a
whole, or often the Neuropathic Symptoms Scores, may not improve, as shown by
several reviews on whether the neuropathy is reversible or not in diabetes later in
this monograph.
A note of nerve conduction velocity and normal values:
In order to complete the perspective on assessment of sensory neuropathy, we would
like the reader to keep a few salient features of the nerve conduction velocity studies.
Nerve conduction (sensory) velocity studies primarily assess the amplitude of the
sensory action potentials and velocities. The amplitudes are basically the summation

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of all the electrical amplitudes of individual axons, which generate them and are
being tested. Thus, any abnormality i.e., decrease in the amplitude is primarily a
function of the number of axons that are available to generate the amplitudes.
Obviously if the neuropathy is of the axonal degenerative type, as a result of diabetes
then it will be primarily reflected as decrease of amplitude from the normal values.
The velocity of electrical impulse that is carried by the axons is dependant upon the
state of myelination - that is either its loss or other structural irregularities. The nerve
fibers that are well myelinated have Ranvier's nodes. These nodes are facilitatory to
the fast transmission of the electrical conduction. As such when there is
demyelination, the velocities drop and the nerve conduction manifests as a
demyelinating type of axonal abnormalities.
Both, the amplitude as well as the velocity can decrease in diabetic neuropathy in
which case a mixed picture of axonal degeneration as well as demyelination emerges
from these studies.
Limitations of NCV:
For the decrements of either velocities and / or amplitudes, a large number of axonal
fibers need to be either dead or demyelinated to get a consistent picture of neural
pathology. The other sensory modalities like vibration or heat and cold could be
detectable earlier than that. Specifically EMG and NCV do not detect the small C
fiber functions which becomes easily detectable by Heat and Cold Perception
Threshold measuring instruments now available in India.
References:
1. Boulton AJM, Kubrusly DB, Bowker JH, et al. TI: Impaired Vibratory Perception
and Diabetic Foot Ulceration Diabet Med 1986; 3:335.
2. Boulton AJM, Mueller MJ. Identifying patients with diabetes mellitus who are at risk
for lower extremity complications. Use of Semmes Weinstein monofilaments. Phys
Ther 1996; 76: 68.
3. Kumar S, Fernando DJ, Veves A, et al. Semmes Weinstein monofilaments: A simple,
effective and inexpensive screening device for identifying diabetic patients at risk of
foot ulceration . Diab Res Clin Pract 1991; 13:63.
4. Young MJ, Bready JL, Veves A, et al. The prediction of diabetic neuropathic foot
ulceration using vibration perception thresholds: A prospective study. Diabetes Care
1994; 17:57.
Note: A table of normal values for the EMG / NCV is given as Appendix obtained
from a reputed neurology institute that is engaged in these studies for last over 15
years. We record our thanks for the gesture of Central India Institute Of Medical
Sciences, 88/2, Bajaj Nagar, Nagpur 440010, and of Dr. G M Taori MD MRCP, Chief
Neurologist and Director and Dr. Leena Vyas M.B.B.S, the electrophysiologist of the
Dept of Neurology.