Detection of cytomegalovirus in upper gastrointestinal biopsies from heart transplant recipients: comparison of light microscopy, immunocytochemistry, in situ hybridisation, and nested PCR.

Abstract

Aim-To establish the diagnostic value of in situ hybridisation and the nested polymerase chain reaction (PCR) in detecting clinically relevant cytomegalovirus (CMV) infection in upper gastrointestinal biopsies from heart transplant patients. Methods-Test sensitivity and specificity for detection of CMV early gene RNA by in situ hybridisation and CMV intermediate early gene by PCR were established and then compared with haematoxylin and eosin (H&E) and immunocytochemical detection of CMV in order to establish the best pathological diagnostic approach. All investigations were carried out on formalin fixed, paraYn embedded tissue. Results-Nested PCR had the highest test sensitivity, followed by in situ hybridisation and immunocytochemistry with the same sensitivity; H&E had the lowest. H&E and immunocytochemistry were the most specific but both had a significant false negative rate which was less of a problem with PCR. However, PCR gave no other diagnostic information, and in situ hybridisation was no better than immunocytochemistry. Both in situ hybridisation and PCR were technically complex and more expensive. Conclusions-H&E and immunocytochemistry represent the best initial screen for CMV and other diseases in upper gastrointestinal biopsies from heart transplant patients. If H&E and immunocytochemistry were negative, nested PCR could significantly increase the diagnostic yield of clinically relevant CMV infection. In situ hybridisation appeared to have no advantages and some drawbacks compared with immunocytochemistry and PCR. (J Clin Pathol 1998;51:807-811)

Cytomegalovirus infection and development of biliary complications after liver transplantation1

“…Standard histology, hematoxylineosin stain, and cytochemistry for formalin-fixed and paraffin-embedded tissue were recommended to screen for CMV (1,19). If these results were negative, PCR increased the diagnostic yield in clinically relevant CMV infection, but in situ hybridization appeared to give no diagnostic advantage compared with immunocytochemistry and PCR (19). CMV culture has been the method of choice for many years in reports of gastrointestinal tract CMV complications (20).…”

New Strategies for Prevention and Therapy of Cytomegalovirus Infection and Disease in Solid-Organ Transplant Recipients

“…Furthermore, the test specificity for allograft biopsy specimens is as high as 100% (286,341,492). Applications for this test include the diagnosis of CMV pneumonitis (113,160,436,493), hep- atitis (286,341) and gastroenteritis (65,308,503). However, the techniques involved are cumbersome.…”

High Prevalence of Viral Genomes and Multiple Viral Infections in the Myocardium of Adults With “Idiopathic” Left Ventricular Dysfunction

“…Polymerase chain reaction (PCR)/reverse transcription PCR (RT-PCR) was performed for the detection of enteroviruses (including coxsackievirus and echovirus), adenovirus, 12 parvovirus B19, 9,13 human herpesvirus type 6, 13 human cytomegalovirus, 14 Epstein-Barr virus, 15 influenza virus A and B, 16 herpes simplex virus 1 and 2, 17 and hepatitis virus C. 18 In addition, DNA was extracted from peripheral blood cells (PBLs) to exclude a systemic infection with PVB19, EBV, and HHV6. As a control for successful extraction of DNA and RNA from heart muscle tissue, oligonucleotide sequences were chosen from the DNA sequence of the GAPDH gene.…”