pain is a diagnostic criterion for Gulf War illness (GWi), chronic fatigue Syndrome (cfS), and fibromyalgia (FM). The physical sign of systemic hyperalgesia (tenderness) was assessed in 920 women who were stratified by 2000 Kansas GWI, 1994 CFS, and 1990 FM criteria. Pressure was applied by dolorimetry at 18 traditional tender points and the average pressure causing pain determined. GWI women were the most tender (2.9 ± 1.6 kg, mean ± SD, n = 70), followed by CFS/FM (3.1 ± 1.4 kg, n = 196), FM (3.9 ± 1.4 kg, n = 56), and CFS (5.8 ± 2.1 kg, n = 170) compared to controls (7.2 ± 2.4 kg, significantly highest by Mann-Whitney tests p < 0.0001, n = 428). Receiver operating characteristics set pressure thresholds of 4.0 kg to define GWI and CFS/FM (specificity 0.85, sensitivities 0.80 and 0.83, respectively), 4.5 kg for FM, and 6.0 kg for CFS. Pain, fatigue, quality of life, and CFS symptoms were equivalent for GWI, CFS/FM and CFS. Dolorimetry correlated with symptoms in GWI but not CFS or FM. Therefore, women with GWI, CFS and FM have systemic hyperalgesia compared to sedentary controls. The physical sign of tenderness may complement the symptoms of the Kansas criteria as a diagnostic criterion for GWI females, and aid in the diagnosis of CFS. Molecular mechanisms of systemic hyperalgesia may provide new insights into the neuropathology and treatments of these nociceptive, interoceptive and fatiguing illnesses.Pain and tenderness (systemic hyperalgesia) are common complaints in Gulf War Illness (GWI) 1-3 , Chronic Fatigue Syndrome (CFS) 4 , and fibromyalgia (FM) 5-8 . The symptoms of myalgia and arthralgia are in the diagnostic criteria for all three conditions, but it is not clear if pain severity can distinguish between them. This ambiguity is of importance because (i) pain was not included as a distinguishing feature in the most recent reconceptualization of CFS as Systemic Exertion Intolerance Disease (SEID) 9 , (ii) fatigue, unrefreshing sleep, and cognitive dysfunction were added to the 2010 6 and 2011 7 American College of Rheumatology criteria for FM, (iii) pain, fatigue, sleep and cognitive dysfunction are defining characteristics of GWI 2,3 . Symptoms alone are insufficient to distinguish between these diseases.An alternative approach to distinguish between GWI, CFS and FM may be to assess the sign of systemic hyperalgesia, the perception of physical discomfort elicited by pressure stimulation 10 . Assessment of tenderness was only required for the 1990 FM criteria 5 . Dolorimetry (algometry) was used to quantify the cutaneous pressure required to induce pain [11][12][13] . This semi-quantitative method provides a more robust and reproducible measurement than traditional tender point counts that are influenced by psychological status 14,15 .A confounding design in studies of hyperalgesia is to compare dolorimetry between tender (e.g. defined using 1990 FM criteria 5 ) versus nontender (general population) groups. Such a study design ensures a floor effect for pressures that cause pain in FM, a ceiling effect ...