A Critical Analysis of Rectal Biopsy to Exclude Hirschsprung's Disease.

Background: The diagnosis of Hirschsprung’s disease (HD) is dependent on the histological study of rectal ganglion cells. Open rectal biopsy is the mainstay that requires general anesthesia (GA) and carries risk of many complications. Rectal punch biopsy has gained wide acceptance and has become the choice as it is simple, safe, having no need of anesthesia, done bedside with a simple biopsy forcep with high degree of accuracy and virtual absence of any complications. Objective: To find out the efficacy of rectal punch biopsy in the diagnosis of Hirschsprung’s disease. Materials and Methods: A cross sectional descriptive observational study was carried out with the histological findings of 60 rectal punch biopsies with corresponding 60 full thickness rectal biopsies from 60 suspected HD patients from March 2018 to February 2020. Sections made from the paraffin blocks were stained with H&E and were examined for the presence or absence of ganglion cells and hypertrophic nerve fibers in the submucosa. Then the relations were studied. These findings were compared with full thickness rectal biopsy. Result: There were 40(66.7%) cases of male and 20(33.3%) cases of female including 26 (43.3%) cases of infants. Out of 60 cases, 10 cases (seven inadequate and three suspicious) of punch biopsy were not compared with full thickness rectal biopsy. Out of 50 cases, 38(63.3%) cases were HD in punch biopsy; among these 36(92.3%) cases were HD and 2(18.2%) cases were Non-HD in full thickness rectal tissue biopsy. The accuracy of punch biopsy was measured by sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV) and efficiency of the test which were 92.3%, 81.8%, 94.7%, 75.0% and 90.0% respectively. Conclusion: The rectal punch biopsy is simple, safe, no need of anesthesia and can be done with a simple biopsy forcep with high degree of accuracy. KYAMC Journal Vol. 13, No. 01, April 2022: 37-40

IntroductionIt is challenging to distinguish between patients with Hirschsprung disease (HD) and patients with other causes of defecation problems based on clinical presentation in patients older than six months. Pathological examination of the rectal biopsy is the gold standard for the diagnosis of HD. The aim of this study was to gain insight into 1) the prevalence and severity of complications following rectal biopsy, 2) the final diagnoses of patients referred for biopsy, and 3) clinical factors associated with HD in patients older than six months. MethodsChildren suspected of HD above the age of six months referred for biopsies were analyzed retrospectively. Severity of complications of rectal suction biopsy (RSB) and full thickness biopsy (FTB) were assessed using Clavien-Madadi (CM) grading. Factors associated with HD were tested using multivariate logistic regression analysis. ResultsFrom 2000 to 2022, 234 children older than six months of age underwent biopsies because of defecation problems (median age of 47.2 months (IQR = 17.2–87.2)). Of these, 130 out of 234 children underwent RSB and 112 out of 234 children underwent FTB. One patient (0.4 %) developed a complication following RSB (CM1-A): fever without evident cause. Two patients (0.9 %) had ongoing rectal bleeding following FTB. One patient did not require an intervention (CM1-A), the other received rectal spongostan to stop the bleeding and erythrocyte transfusion (CM2). The most frequent final diagnoses were functional constipation (n = 179, 76.5 %), HD (n = 25, 10.6 %) and food intolerance (n = 5, 2.1 %). Associated factors for HD were distended abdomen (OR 5.41, CI 2.05–14.31), vomiting (OR 4.14, CI 1.64–11.85) and no abdominal pain (OR 0.14, CI 0.03–0.65) (model R2 = 0.278). ConclusionIn children older than six months suspected of HD, presenting with distended abdomen, vomiting and no abdominal pain, we advise obtaining a rectal biopsy in case other causes of constipation are ruled out, because a rectal biopsy is a safe procedure with only minor complications. The most common diagnosis in patients older than six months of age referred for rectal biopsy was functional constipation. Level of EvidenceLevel III.

C.W.WongC.T.LauP.H.ChungThe value of the 24-h delayed abdominal radiograph of barium enema in the diagnosis of Hirschsprung's diseasePediatr Surg Int31120151115

The objective of this study is to determine the diagnostic value of the 24-h delayed film for Hirschsprung's disease (HD). Other features of the barium enema were also examined, in particular the correlation between the radiological transition zone (TZ) and the final pathology. All patients with suspected HD from 2003 to 2013 who had undergone barium enema and rectal biopsy were reviewed retrospectively to study the correlation between radiological features of barium enema with the final diagnosis as well as severity. A total of 182 patients were admitted for suspected HD during the study period, of which 82 had both investigations done. 68 patients had radiological features suggestive of the disease and ultimately, 12 patients had the disease confirmed with rectal biopsy. Among those without radiological features of HD, 2 patients were found to have the disease. Thus, the sensitivity of the 24-h delayed film was 85.7% and the specificity was 17.6%. The positive predictive value (PPV) of this test was 20.6% and the negative predictive value (NPV) was 85.7%. Regarding the level of TZ, it was not detected in the barium enema in 7 (50%) out of the 14 patients. For those with the presence of TZ, 6 (85.7%) of them correlated well with the intra-operative findings and 4 (57.1%) of them correlated well with the final histology. The 24-h delayed film of barium enema has a high NPV and is useful to rule out HD. However, rectal biopsy is still suggested for disease confirmation given its low PPV. Lastly, once present, the level of radiological TZ is also a useful predictor for the actual disease involvement.

The presenting symptoms of patients with Hirschsprung's disease (HD) are a failure to pass meconium, abdominal distension, and bilious vomiting. The gold standard diagnosis is a rectal biopsy to confirm aganglionosis. The aim of our study was to describe the diagnostic pathway of Hirschsprung's disease at our institution and document the indication for a rectal biopsy. We have performed a prospective collection of all patients who underwent a rectal biopsy to exclude HD from December 2022 until September 2023 including. The following data were collected: patient's age, presenting symptoms, type of biopsy, failure rate, complications, and histopathological results. We identified 33 patients who underwent 34 rectal biopsies at 0.6 years of age. A total of 17 patients had a rectal suction biopsy (RSB), and 17 patients underwent a partial thickness under general anaesthesia (GA). 1/17 (6%) patients had an inconclusive RSB and subsequently underwent a biopsy under GA. Constipation and chronic abdominal distension plus vomiting were the most common presenting symptoms throughout all ages. Five patients (15%) had a rectal biopsy that was positive for HD. A protocolised approach to the assessment of infants and children with suspected HD ensures the appropriate utilisation of invasive procedures such as biopsy.

Rectal biopsy is considered the gold standard for the diagnosis of Hirschsprung's disease. The aim of this study was to evaluate the outcome of rectal biopsies performed in our institution, and to determine whether we are performing an adequate number of biopsies in patients presenting with features suggestive of this disease. A retrospective analysis was conducted of patients who underwent rectal biopsy to exclude Hirschsprung's disease over a seven year period between 2000 and 2006. The histological diagnosis of Hirschsprung's disease was made using haematoxylin and eosin as well as acetylcholinesterase on frozen section. Patients were grouped into three age categories: neonates (group A), infants (group B) and those over 1 year of age (group C). The results of the biopsies were compared between groups. A total of 668 patients underwent rectal biopsy. 18 samples were insufficient. Based on the histological studies of 650 suitable samples, 73 (11%) were positive and 577 (89%) were negative for Hirschsprung's disease. Of the 73 positive biopsies, 34 (47%) were from group A, 20 (27%) from group B and 19 (26%) from group C. The percentage of positive biopsies was much higher in group A with 29% (34 out of 118) compared to group B with 15% (20 out of 135) and group C with 5% (19 out of 395). Three complications of minor rectal bleeding occurred. With 3 complications and 18 insufficient samples out of 668, rectal biopsy is a safe procedure and remains the gold standard for the diagnosis of Hirschsprung's disease, despite the large number of negative biopsies. Contrary to some reports in the literature which question the need for rectal biopsy in those presenting after the neonatal period, 53% of our positive diagnoses were made in children presenting after this period, with 19 positive biopsies out of 395 (5%) performed in children above the age of 1 year.

The gold standard for the diagnosis of Hirschsprung's disease (HD) is a rectal biopsy. The sample may be obtained using a transanal excisional biopsy (TEB) or suction technique. Rectal suction biopsy (RSB) is not a standard procedure in Latin-America. To evaluate the current practice in rectal biopsy for HD among pediatric surgeons. We distributed an online questionnaire among Latin-American pediatric surgeons. One hundred forty-nine pediatric surgeons from 15 countries completed the anonymous survey (71.4% of Latin-American countries), grouped into 81.9% pediatric surgeons; 8.9% pediatric colorectal surgeons, 8.9% trainees, and 1 pediatric colorectal surgeon fellow. 50.4% reported less than 5 new patients with HD per year, 36.2% 5-10 new cases, and 13.4% more than 10. Only 14.1% of surgeons have access to perform a RSB in the diagnostic work-up of patients with suspected HD, 90% in our study perform an TEB under general anesthesia. When we ask if they could perform both procedures in babies up to 6months, 52.3% indicate that they prefer an RSB, and for patients older than 6months, 35.4% favor an RSB. Regarding the number of samples obtained performing an TEB, 30.9% get one biopsy, 29.5% two biopsies, and 39.6% three or more samples. Surgeons obtained the most proximal biopsy at a median of 2.3cm (range 1-4cm) above the pectinate line. 67.8% of surgeons prescribed antibiotic prophylaxis. Overall, 16.1% experienced complications, including rectal blood loss (n = 18), and rectal perforation (n = 3). The most frequently used staining methods for rectal biopsies are hematoxylin/eosin (87%), calretinin (56, 8%), and acetylcholinesterase (21.9%). In Latin-America, the accessibility for RSB is limited only 18 out of 149 surgeons have access to rectal suction tool. There is no consensus regarding sample number, site of proximal biopsy, and antibiotics use. The complications associated with the procedure seems to be less than reported with RSB. Therefore, we should standardize this common surgical practice and establish universal guidelines for rectal biopsy procedure (RBP).

Diagnosing Hirschsprung's disease: Increasing the odds of a positive rectal biopsy result

The diagnosis of Hirschsprung's disease is usually made in neonates but often considered in older infants and children with constipation: these children may be referred for barium enema. Since it is widely accepted that a normal barium enema does not exclude Hirschsprung's disease, some children, after a normal enema, undergo more invasive procedures such as rectal manometry or biopsy. Our study asked how frequently a diagnosis of Hirschsprung's disease was made by biopsy or manometry in children who had normal barium enema. We reviewed the medical records and barium enemas of 54 patients older than 28 days with constipation or difficulty passing stool who had a barium enema followed by manometry and/or biopsy. Forty-eight patients had normal enemas: 24 of those patients had biopsies, 16 had manometry, and 8 both manometry and biopsy. Only 1 had manometry suggestive of Hirschsprung's disease, confirmed by biopsy. Six patients had abnormal enemas. Five had biopsy and manometry compatible with Hirschsprung's disease; one had a normal biopsy and manometry study. The barium enema is a good initial screening test for Hirschsprung's disease in severely constipated children since it correlates well with manometry and biopsy. The enema is particularly useful in centers without easy access to pediatric gastroenterology services, and a normal enema in this setting allows the continuation of medical therapy with further evaluation only if there is a lack of response. An abnormal enema, however, requires referral to a facility equipped to perform confirmatory manometry or biopsy.

Objective To explore the diagnostic value of barium enema film for Hirschsprung’s disease (HD) and examine the correlation between radiological transition zone (TZ) and final pathology. Methods For this single-center cross-sectional study, a total of 56 patients with suspected HD undergoing barium enema and primary pull-through operations from January 2013 to January 2014 were reviewed. According to barium enema, full-thickness bowel wall biopsies were performed before and after 5 cm of transitional zone. And the gold standard was detecting ganglion cell by PGP9.5 staining. If aganglionosis was considered positive whereas ganglionosis negative. The diagnostic parameters of sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of barium enema in the range of HD were analyzed by statistical software. Results Their average age was 13.7 months. The sensitivity of barium enema film was 86.9% and specificity 94.1%. The value of PPV was 94.6% and that of NPV 85.7%. Youden index was 81%. Consistency checking Kappa value was 0.804 (P<0.001). The diagnostic coincidence rate was 77.8% in short-segment HD, 88.9% in normal-segment HD and 66.7% in long-segment HD. Conclusions With a high diagnostic predictive value for HD, barium enema film is useful for confirming the involvement extent of aganglionosis bowel. Key words: Hirschsprung’s disease; Barium enema; Child

Radiologist performance in the interpretation of contrast enemas performed for Hirschsprung's disease in children >1 year of age

Hirschsprung disease (HD) is defined as the absence of ganglion cells in the Meissner and Auerbach plexuses. Diagnosis depends on demonstrating the absence of ganglion cells in rectal biopsy specimens. Rectal suction biopsy is widely employed as the method of choice in obtaining such specimens. Classically, the diagnosis was made until the 1990s by using the Multipurpose Suction Biopsy Kit, or Rubin Tube. This device was replaced by the Model SBT-100 Suction Biopsy Kit as the exclusive device used to procure rectal tissue. Because the suction devices are known to occasionally yield tissue that is insufficient, the present study was undertaken to determine the efficacy of using this technique to make or exclude the diagnosis of HD. The last 50 biopsy sessions using the Multipurpose Suction Biopsy Kit and the first 46 sessions using the Model SBT-100 were included for review. Both groups had similar yields (24%) of biopsy sessions with insufficient tissue to allow meaningful interpretation. The predictive value of rectal suction biopsy in excluding HD at the first biopsy session was 65%. We conclude that HD can be excluded with a single rectal suction biopsy 65% of the time. A second biopsy session will exclude the diagnosis in an additional 11% of patients. Both devices yield biopsies of comparable quality and are equally useful in excluding the diagnoses of HD.

Background: The golden standard of Hirschsprung's disease diagnosis is through a histopathological examination of the aganglionic site. The significance of serum acetylcholinesterase (AChE) immunohistochemistry for diagnosing Hirschsprung's disease has been widely accepted. However, the study of serum AChE levels in Hirschsprung's disease patients is still scarce. We aimed to examine the serum levels of AChE in individuals with Hirschsprung's disease. Methods: An analytical observational study with a cross sectional design was conducted on 29 patients with Hirschsprung's disease. We divided the patients into two groups: 14 in Hirschsprung's disease group and 15 in the control group. Serum AChE level was measured using the enzyme-linked immunosorbent assay (ELISA) method. The optimal cut-off value for detecting Hirschsprung’s disease was determined using the ROC analysis. Results: The prevalence of Hirschsprung was higher in males than in females. The average AChE level in the Hirschsprung group was 95.89 ± 51.11 Units/mL, while the average level of AChE in the control group was 44.45 ± 33.40 Units/mL. The optimal cut-off value was 46.615 Units/mL. We found that this study's sensitivity, specificity, positive predictive value, negative predictive value, and accuracy values ​​were 83.3%, 70.6%, 66.7%, 85.7%, and 75.9%, respectively. There was a significant relationship between serum AChE levels and Hirschsprung's disease (p=0.004). Conclusion: The AChE levels in blood serum can be used as an alternative diagnostic parameter for Hirschsprung's disease. Patients with Hirschsprung's disease had higher serum AChE levels than patients without Hirschsprung's disease.

The heterogeneity of rectal biopsy techniques has encouraged us to search for a surgical and pathological standardisation of this diagnostic technique to exclude Hirschsprung's disease. The different amounts of information on the anatomopathology report prompted us to compile a template for the anatomopathology report for diagnostic rectal biopsies for surgical colleagues and pathologists working on Hirschsprung's disease. We gathered the anonymous biopsy information and its pathology information from five hospitals for all patients in which rectal biopsies were taken to diagnose Hirschsprung's disease over two years (2020-2021). Of the 82 biopsies, 20 suction (24.4%), 31 punch (37.8%) and 31 open biopsies (37.8%) were taken. Of all biopsies, 69 were conclusive (84.2%), 13 were not (15.8%). In the suction biopsy group, 60% were conclusive and 40% were not; for punch biopsy, the values were 87% and 13%, respectively and for open biopsy, 97% and 3%. Inconclusive results were due to insufficient submucosa in 6/8 suction biopsies, 4/4 punch biopsies and 0/1 open biopsies. An insufficient amount of submucosa was the reason for an inconclusive result in 6/20 cases (30%) after suction biopsy, 4/31 (12.9%) cases after punch biopsy and 0 cases (0%) after open biopsy. We had one case with major postoperative bleeding post suction biopsy; there were no further adverse effects after biopsy. Diagnostic rectal biopsies in children are safe. Non-surgical biopsies are more likely to give inconclusive results due to smaller amounts of submucosa present in the specimen. Open biopsies are especially useful when previous non-surgical biopsies are inconclusive. An experienced pathologist is a key factor for the result. The anatomopathology report should specify the different layers present in the specimen, the presence of ganglion cells and hypertrophic nerve fibres, their description and a conclusion.

Hirschsprung disease (HD) manifests as a developmental anomaly affecting the enteric nervous system, where there is an absence of ganglion cells in the lower part of the intestine. This deficiency leads to functional blockages within the intestines. HD is usually confirmed or ruled out through rectal biopsy. The identification of any ganglion cells through hematoxylin and eosin (H&E) staining rules out HD. If ganglion cells are absent, further staining with acetylcholine-esterase (AChE) histochemistry or calretinin immunohistochemistry (IHC) forms part of the standard procedure for determining a diagnosis of HD. In 2017, our Institute of Pathology at University Hospital of Heidelberg changed our HD diagnostic procedure from AChE histochemistry to calretinin IHC. In this paper, we report the impact of the diagnostic procedure change on surgical HD therapy procedures and on the clinical outcome of HD patients. We conducted a retrospective review of the diagnostic procedures, clinical data, and postoperative progress of 29 patients who underwent surgical treatment for HD in the Department of Pediatric Surgery, University of Heidelberg, between 2012 and 2021. The patient sample was divided into two groups, each covering a treatment period of 5 years. In 2012-2016, HD diagnosis was performed exclusively using AChE histochemistry (AChE group, n = 17). In 2017-2021, HD diagnosis was performed exclusively using calretinin IHC (CR group, n = 12). There were no significant differences between the groups in sex distribution, weeks of gestation, birth weight, length of the aganglionic segment, or associated congenital anomalies. Almost half of the children in the AChE group, twice as many as in the CR group, required an enterostomy before transanal endorectal pull-through procedure (TERPT). In the AChE group, 4 patients (23.5%) required repeat bowel sampling to confirm the diagnosis. Compared to the AChE group, more children in the CR group suffered from constipation post TERPT. Elevated AChE expression is linked to hypertrophied extrinsic cholinergic nerve fibers in the aganglionic segment in the majority of patients with HD. The manifestation of increased AChE expression develops over time. Therefore, in neonatal patients with HD, especially those in the first 3 weeks of life, an increase in AChE reaction is not detected. Calretinin IHC reliably identifies the presence or absence of ganglion cells and offers multiple benefits over AChE histochemistry. These include the ability to perform the test on paraffin-embedded tissue sections, a straightforward staining pattern, a clear binary interpretation (negative or positive), cost-effectiveness, and utility regardless of patient age. The ability of calretinin IHC to diagnose HD early and time-independently prevented repeated intestinal biopsies in our patient population and allowed us to perform a one-stage TERPT in the first months of life, reducing the number of enterostomies and restoring colonic continuity early. Patients undergoing transanal pull-through under the age of 3 months require a close follow-up to detect cases with bowel movement problems.