fibrosis-type symptoms and a false-positive sweat test SMN Brown 1 + R Buchdahl 1,2 1 Department of Paediatrics, Hillingdon Hospital NHSTrust, London, UK 2 Department of Paediatric Respiratory Medicine, Royal Brompton and Harefield NHSTrust, London, UK Correspondence to: Sarah Brown. E-mail: sarahbrownmay@doctors.org.uk Introduction A six-month-old Caucasian boy was referred by his general practitioner with concerns regarding poor weight gain and poor feeding. He was the second child of unrelated parents. His mother was Italian and his father English. His older brother, aged 5 years, was known to be egg allergic but well in other respects. There was no other family his- tory of note. He was born following a normal preg- nancy and there were no initial neonatal concerns. Birth weight was 3.86 kg (75th centile). He was breast-fed for the first four months, after which he was fed on standard formula. From 14 weeks of age his weight started to falter from the 75th to the 9th centile. He hadonly receivedhis first immuniz- ation at two months of age due to parental con- cerns regarding his health. There had been no developmental concerns. On his first reviewhe had appeared quite irrita- ble on feeding and the question of cows milk pro- tein intolerance was raised. His feed was therefore changed to a hydrolysed formula and a dietitian reviewed him. On reviewthree weeks later he was found to be chesty and wheezy, but with no chest deformity. He was pale and thin. His stools were noted to be fatty, loose and offensive and his weight gain had not significantly improved. From his initial blood tests he was noted to have a raised alanine aminotransferase (ALT) concentration of 73 U/L (reference range 045 U/L) and a border- line low neutrophil count of 1.9 10 9 /L (reference range 1.58 10 9 /L). The results of other initial investigations, performed as part of a failure-to- thrive screen, were unremarkable, including immunoglobulin E (IgE), radioallergosorbent test (RAST) to cows milk protein, thyroid function tests, urine andfaecal microbiology, coeliac screen, faecal reducing substances and amino and organic acids. Abdominal ultrasound scan was also nor- mal. Given the concerns regarding his poor weight gain, loose offensive stools and malabsorption, a sweat test and faecal elastase level were requested. His sweat test was reported as positive (chlo- ride 75 mmol/L). Sodium levels were not measured. Sweat volume of at least 30 L was collected using the Macroduct sweat collection system. Faecal elastase level was significantly low (<15 IU). His chest X ray was normal. A diagnosis was made of cystic fibrosis (CF) with pancreatic insufficiency. His parents were counselled regard- ing the diagnosis and he was started on prophylac- tic amoxicillin and clavulanic acid, as well as pancreatic enzyme and vitamin supplementation. Genotyping was requested and he was referred to a tertiary centre for joint care. Over the next weeks he started to gain weight, although not as much as would have been expected. His stools were less oily. Genotyping failed to show any CF mutations both on routine analysis and on extended testing. His parents became increasingly concerned that the diagnosis was incorrect. Their concerns were initially raisedas they felt he didnot taste salty, but were compounded by the fact that a mutation was not isolated. A repeat sweat test was sent and reported as negative (chloride 5 mmol/L). A third sweat test was also negative (8 mmol/L). Repeat faecal elastase level was still <15 IU/g, confirming pancreatic insufficiency. Following these two negative sweat tests an alternative diagnosis was sought, in particular Shwachman-Diamond syndrome (SDS). Genotyp- ing was requested and he was found to be homozygous for 258+2T>2C, the genetic mutation for SDS. His parents were both heterozygous. A final diagnosis was therefore made of SDS and he was referred to a tertiary centre for further man- agement. Why was the initial sweat test positive? The sweat tests were performed using the Macrod- uct sweat collection system. At least 30 L of sweat DECLARATIONS Competing interests None declared Funding None Ethical approval Not applicable Guarantor SMNB Contributorship Both authors contributed equally Acknowledgements None SUPPLEMENT J R Soc Med 2008: 101: S39S43. DOI 10.1258/jrsm.2008.s18009 S39 was collected for each test. Chloride level only was analysed by our laboratory following national guidelines. 1 Sodium levels were not analysed. The childs arm was cleaned before each test with dis- tilled water and all other quality controls were met. The laboratory commented that before the second and third sweat tests the mother had cleanedthe childs armwith holy water fromLour- des. They therefore queried whether she had also cleaned the child with the same water before the initial sweat test, and whether this water had a high chloride level, thus affecting the results. A sample of the water was taken for analysis and found not to contain any chloride ions, therefore disproving this theory. The laboratory felt that the only possible area of error was if the childs arm was not cleaned adequately on the first test. They were unable to provide an alternative explanation. There is a well-recognized list of causes of false- positive sweat test results in the literature. 2 How- ever, the evidence for these is limited. The most important of these causes are technical failure and operator error, eczema and other skin conditions where it is often difficult to obtain enough sweat, and underlying blood electrolyte abnormalities which may cause misleading results. To our knowledge there are no cases reportedof SDS itself causing a false-positive sweat test, although con- ceivably any underlying illness which causes mal- nutrition may cause misleadingly positive results. 3 This particular case scenario is clearly not par- ticularly common and hopefully with the advent of national neonatal screening for CF the situation will become even less likely. It does, however, highlight some important points, including remembering other causes of malabsorption and failure to thrive in children. This case serves as a reminder to always repeat a sweat test if there is any doubt regarding the diagnosis. In this particu- lar case, the presence of pancreatic insufficiency and convincing clinical features, along with a posi- tive sweat test, resulted in a diagnosis of CF being confirmed on the basis of one sweat test. Recom- mended practice is to repeat the sweat test up to three times in an attempt to eliminate error. 4 While it is unusual not to find a CF genotype in Caucasian patients, this family were initially reas- suredthat the failure to isolate a genotype is some- times encountered in patients with CF. While this is certainly not incorrect information, it serves as a reminder that even with convincing clinical fea- tures, if a genotype has not been isolated it is worth reconsidering whether the diagnosis is indeed correct. Finally, this is a valuable lesson in taking heed of parental concerns. They may not always be cor- rect and may have great difficulty in accepting a diagnosis such as CF or any other life-limiting condition, but their concerns should always be listened to. Shwachman-Diamond syndrome SDS is an autosomal recessive condition affecting between 1:100,000 and 1:200,000 live births. The majority of patients have compound heterozygous mutations of the Shwachman-Diamond-Bodian syndrome (SBDS) gene on chromosome 7 (7q11). This gene was identified in 2002 and is thought to encode a protein of 250 amino acids. There is an adjacent pseudogene on 7q11 which appears to recombine with the SDBS gene during meiosis, thus making it dysfunctional. 5 SDS was first described in the early 1960s by Nezelof and Watchi, initially as a syndrome encompassing pan- creatic exocrine insufficiency and neutropenia. 6 Shwachman et al. further described the condition as pancreatic insufficiency with bone marrowdys- function in 1964. 7 Skeletal dysplasias were observed to be associated with SDS in the late 1960s. 8 Subsequently other organs have been noted to be involved in children with SDS, with a wide variety of phenotypes. SDS is the second most common inherited cause of pancreatic exocrine insufficiency after CF, and should therefore be included in the differential diagnosis of children presenting with malabsorp- tion or CF-type symptoms. Usually the two con- ditions can be differentiated on the basis of a sweat test, and this should form part of the initial inves- tigations. Other rarer causes of pancreatic exocrine insufficiency include severe malnutrition, Pearson syndrome, Johanson-Blizzard syndrome and Jeune syndrome. Clinical features Exocrine pancreatic insufficiency Pancreatic insufficiency in SDS presents with stea- torrhoea and malabsorption in the first months of life. These children have poor growth and often initially present due to failure to thrive. A pro- portion of children will still be pancreatic suf- ficient, as it is necessary to lose over 98%pancreatic exocrine function before clinical malabsorption becomes apparent. Children may also be deficient in the fat-soluble vitamins A, D, E and K due to fat malabsorption. Journal of the Royal Society of Medicine J R Soc Med 2008: 101: S39S43. DOI 10.1258/jrsm.2008.s18009 S40 Unlike CF, pancreatic exocrine function in SDS improves over time in 4060% of patients. 9 This can make diagnosis in older children more diffi- cult, as they may appear pancreatic sufficient. Since the isolation of the SDS gene and successful molecular genetics, it is now possible to make a diagnosis of SDS in most children. The underlying pathophysiology of pancreatic insufficiency in SDS is thought to be due to replacement of the pancreatic exocrine acini with fat. This is different to CF where pancreatic insuf- ficiency is due to obstruction by thickened secre- tions. In SDS there is preservation of the endocrine activity as the islets of Langerhans and ductal structure are spared. Fatty pancreatic infiltration can be seen on imaging such as ultrasound scan or abdominal computed tomography. 10 Pancreatic insufficiency and fat malabsorption can be assessed by a number of means. Direct pancreatic stimulation is considered the gold stan- dard, but is rarely used due to the invasiveness of the test. Three-day faecal fat measurement is also considered by many to be a definitive test. How- ever, this is unpleasant for the family and is becoming less available as a laboratory investi- gation. Spot measurements of excessive numbers of fat globules in stool can be useful as a basic screening test where other investigations are not available, but this can only provide limited infor- mation. Many centres rely on faecal elastase measure- ment as an assessment of exocrine sufficiency. This has been shown overall to be a reliable marker of pancreatic function. However, low lev- els can also be seen in patients with malabsorption from a primary intestinal cause, rather than pan- creatic insufficiency. 11 Serumpancreatic enzymes (iso-amylase and trypsinogen) should also be assayed and will be low in SDS. Similarly, immune reactive trypsinogen (IRT) levels will be normal or low. 12 This is as a contrast to CF, where IRT levels will be high due to ductal obstruction. 13 In normal subjects, serum trypsinogen levels are present at birth and through infancy. This is in contrast to serum iso-amylase levels, which are naturally low at birth and in infancy, making interpretation of levels in children <3 years old more difficult. Serum trypsinogen levels will rise with improving pancreatic function in some patients, with approximately 20% having normal levels by the age of 34. 14 It is therefore important to stress that apparently normal pancreatic func- tion suspected by the absence of steatorrhoea or normal pancreatic enzyme levels does not exclude a diagnosis of SDS. As well as infiltration of the pancreas with fat, children may also develop fatty liver. Around 60% of children will have raised liver transaminases and hepatomegaly, although generally no other features of hepatic dysfunction. Over time transaminase concentrations and liver size return to normal. 15 Haematological abnormalities A number of different haematological abnormali- ties have been described in SDS. It is generally agreed that the development of haematological abnormalities in SDS is related to low numbers of CD34+ progenitor cells and a faulty marrow microenvironment. 16 There does not appear to be a genotypephenotype correlation for haematologi- cal disorders in patients with SDS. 17 The most common abnormality seen is neutropenia, which can be continuous but more commonly varies over time. 18 The neutrophils that are present are usually functionally impaired, thus exacerbating suscepti- bility to bacterial infections. SDS neutrophils retain the ability to migrate to areas of infections, but lack the ability to localize and kill pathogens. Patients are susceptible to a wide variety of infections, including pneumonia, which may initially confuse the clinical picture with CF. Particular pathogens which SDS patients are susceptible to include Sta- phylococcus aureus, Haemophilus influenzae and Gram-negative pathogens including Pseu- domonas species. Many will require prophylactic antibiotic therapy to try to prevent the develop- ment of bacterial infections. 19 Granulocyte colony- stimulating factor has been used to stimulate neutrophil levels in some patients, although con- cern has been raised that this therapy could increase the risk of acute myeloid leukaemia (AML). 18 Patients with SDS should be monitored by rou- tine blood testing and bone marrow assessment at diagnosis andthroughout life for bone marrow failure, myelodysplasia and malignant change, in particular AML. Around 2033% of patients will develop myelodysplasia, with 1225% developing leukaemia. 20 Treatment of myelodysplasia or AML with chemotherapy is often unsuccessful. Bone marrow transplantation is seen as an option in some patients with severe haematological abnormalities, with varying success. 20 SDS patients may have isolated cytopenias other than neutropenia, including anaemia and thrombocytopenia, and a significant proportion develop aplastic anaemia. Shwachman-Diamond syndrome in a child presenting with cystic fibrosis-type symptoms J R Soc Med 2008: 101: S39S43. DOI 10.1258/jrsm.2008.s18009 S41 Skeletal and growth abnormalities Most patients with SDS are growth restricted, and this is often present at birth. This suggests that not only does malabsorption have a significant impact on growth and development, but that there is also an inherent abnormality of growth in these patients. Most patients will remain small through- out their lives, with associated pubertal delay. 21 There are a number of skeletal abnormalities present in patients with SDS, but their pathogen- esis is unclear. Metaphyseal dysostosis is com- monly found (4080% of patients), particularly in the femoral head, hips, spine andknees. Rib abnor- malities occur in around 50% of patients, with chostochondral thickening of shortened ribs. Abnormalities of the digits have also been described. 13 Patients with SDS are also at risk of defects of bone mineralization due to deficiencies in vitamin D secondary to malabsorption. Psychological abnormalities Since the first description of SDS it has become clear that many children with SDS have learning difficul- ties and developmental delay, as well as some behavioural problems. Studies have suggested that these are of a neurological rather than social aeti- ology, and are part of the disease process. 22 Other abnormalities Since the original description of SDS, a wide var- iety of other abnormalities and organ involve- ments have been described. These include dental abnormalities, dermatological conditions such as severe eczema and ichthyosis, facial abnormalities, myocardial fibrosis, renal calculi and renal tract abnormalities. 23 Management of children with Shwachman-Diamond syndrome As with all chronic conditions, a multi-disciplinary approach to management is appropriate. Children require pancreatic enzyme and fat-soluble vitamin supplementation, with particular attention given to their nutritional status, growth and develop- ment. Involvement of a paediatric dietician is para- mount, as is the involvement of a paediatric gastroenterologist. Many patients develop improving pancreatic function, and enzyme sup- plementation requirements may therefore reduce over time. 24 Routine monitoring of the patients haemato- logical status is required at diagnosis and through- out life. This includes bone marrow assessment. Patients with febrile neutropenia should receive intensive broad-spectrum antibiotic cover. Those with cytopenias may require blood and platelet transfusions. Appropriate specialities should be involved in the management of other clinical manifestations of the disease. Life expectancy of patients with SDS is expected to be >35 years. 13 However, those with significant haematological abnormalities, including AML, have significant morbidity and mortality and sub- sequently reduced life expectancy. References 1 Green A, Kirk JGuidelines Development Group. Guidelines for the performance of the sweat test for the diagnosis of cystic fibrosis. Ann Clin Biochem 2007;44:2534 2 Dinwiddie R. Cystic Fibrosis. The diagnosis and management of paediatric respiratory disorders. London: Churchill Livingstone; 1990. p. 184 3 Yigit H, Selimoglu MA, Altinkaynak S. Sweat test results in children with primary protein-energy malnutrition. J Pediatr Gastroenterol Nutr 2003;37:2425 4 Wallis C. Diagnosing cystic fibrosis: blood, sweat and tears. Arch Dis Child 1997;76:858 5 Woloszynek JR, Rothbaum RJ, Rawls AS, et al. Mutations of the SBDS gene are present in most patients with Shwachman-Diamond syndrome. Blood 2004;104:358890 6 Nezelof C, Watchi M. Lhypoplasie congenitale lipomateuse du pancreas exocrine chez lenfant (Deux observations et revue de la literature). Arch Franc de pediat 1961;18:113572 7 Shwachman H, Diamond LK, Oski FA, et al. The syndrome of pancreatic insufficiency and bone marrow dysfunction. J Pediatr 1964;65:64563 8 Pringle EM, Young WF, Haworth EM. Syndrome of pancreatic insufficiency, blood dyscrasia and metaphyseal dysplasia. Proc R Soc Med 1968;61:7767 9 Cano DA, Hebrok M, Zenker M. Pancreatic development and disease. Gastroenterology 2007;132:74562 10 Stormon MO, Durie PR. Pathophysiologic basis of exocrine pancreatic dysfunction in childhood. J Pediatr Gastroenterol Nutr 2002;35:821 11 Bebarry S, Ellis L, Corey M, et al. How useful is fecal pancreatic elastase 1 as a marker of exocrine pancreatic disease? J Pediatr 2002;141:8490 12 Ip WF, Ellis L, Beharry S, et al. Serum pancreatic enzymes in patients with Shwachman-Diamond syndrome. Annual meeting of the North American society for pediatric gastroenterology and nutrition; October 1999 13 Hall GW, Dale P, Dodge JA. Shwachman-Diamond syndrome: UK perspective. Arch Dis Child 2006;91:5214 14 Ip WF, Dupuis A, Ellis L, et al. Serum pancreatic enzymes define the pancreatic phenotype in patients with Shwachman-Diamond syndrome. J Pediatr 2002;141:25965 15 Mack DR, Forstner GG, Wilschanski M, et al. Shwachman syndrome: exocrine pancreatic dysfunction and variable phenotypic expression. Gastroenterology 1996;111:1593602 16 Dror Y, Freedman M. Shwachman-Diamond syndrome: An inherited preleukemic bone marrow failure disorder with aberrant hematopoietic progenitors and faulty marrow microenvironment. Blood 1999;94:304854 Journal of the Royal Society of Medicine J R Soc Med 2008: 101: S39S43. DOI 10.1258/jrsm.2008.s18009 S42 17 Kuijpers TW, Alders M, Tool ATJ, et al. Hematologic abnormalities in Shwachman-Diamond syndrome: lack of genotype-phenotype relationship. Blood 2005;106:35661 18 Dror Y, Freedman MH. Shwachman-Diamond syndrome. A review. Br J Haematol 2002;118:70113 19 Grinspan Z, Pikora C. Infections in patients with Shwachman-Diamond syndrome. J Ped Infect Dis 2005;24:17981 20 Hsu JW, Vogelsang G, Jones RJ, et al. Bone marrow transplantation in Shwachman-Diamond syndrome. Bone Marrow Transplant 2002;30:2558 21 Cipolli M. Shwachman-Diamond syndrome: Clinical phenotypes. Pancreatology 2001;1:5438 22 Kent A, Murphy GH, Milla P. Psychological characteristics of children with Shwachman syndrome. Arch Dis Child 1990;65:134952 23 Rothbaum R, Perrault J, Vlachos A, et al. Shwachman-Diamond syndrome: Report from an international conference. J Pediatr 2002;141:26670 24 Cipolli M, DOrazio C, Delmarco A, et al. Shwachmans syndrome: Pathomorphosis and long-term outcome. J Pediatr Gastroenterol Nutr 1999;29:26572 Shwachman-Diamond syndrome in a child presenting with cystic fibrosis-type symptoms J R Soc Med 2008: 101: S39S43. DOI 10.1258/jrsm.2008.s18009 S43