New Approaches for the Diagnosis of Lymphoma and Multicentric Castleman Disease

To address the need for new approaches to diagnose lymphoma and multicentric Castleman disease (MCD), investigators have developed an assay based on nucleic acid measurement that has the potential to be delivered at the point of clinical care. It has demonstrated the ability to detect Epstein-Barr virus (EBV) from fine needle aspirations (FNA) and from tumor cells using the Tiny Isothermal Nucleic acid Amplification System (TINY) device, employing primers for EBV-encoded small RNAs (EBER). The method takes advantage of the now well-established finding that Kaposi sarcoma herpesvirus (KSHV) is nearly entirely sensitive (though not specific) for the diagnosis of MCD.

• Lymph Node or Core Needle Biopsy (Procedure): DNA isolation to be tested by nucleic acid amplification using the TINY device
• Fine Needle Aspiration (Procedure): The sample will be eluted into two tubes, one for Gene Xpert testing and one for storage for EBV and KSHV testing by qPCR and TINY.
• Tiny Isothermal Nucleic acid Amplification System (TINY) (Diagnostic test): Tiny Isothermal Nucleic acid Amplification System (TINY)

Primary outcomes

• Number of patients with diagnoses [Enrollment]: Proportion of patients diagnosed with each of the following by histopathology and immunohistochemistry: tuberculosis, lymphoma, MCD, other diagnosis, or non-diagnostic.

Secondary outcomes

• Baseline clinical and laboratory characteristics [Enrollment]: HIV viral load (the number of copies of HIV RNA per milliliter of blood plasma, copies/mL)
• Baseline clinical and laboratory characteristics [Enrollment]: CD4 count
• Baseline clinical and laboratory characteristics [Enrollment]: complete blood count
• Baseline clinical and laboratory characteristics [Enrollment]: creatinine and liver panel
• Baseline clinical and laboratory characteristics [Enrollment]: plasma storage
• Baseline clinical and laboratory characteristics [Enrollment]: buffy coat storage
• Diagnostic performance of KSHV DNA quantification using TINY [Enrollment]: Using lymph node fine needle aspiration (FNA) and tissue biopsy samples using TINY for the diagnosis of KSHV
• Diagnostic performance of EBV DNA quantification using TINY [Enrollment]: Using lymph node fine needle aspiration (FNA) and tissue biopsy using TINY for the diagnosis of EBV
• Performance of KSHV DNA quantification using TINY [Enrollment]: Using a peripheral blood sample using TINY for the diagnosis of KSHV
• Performance of EBV DNA quantification using TINY [Enrollment]: Using peripheral blood sample using TINY for the diagnosis of EBV

Sex: All, ages: 1 Year to any

Inclusion Criteria:

* Age ≥1 year old and body weight of ≥ 10 kg.
* HIV-infected.
* Unexplained lymphadenopathy with at least one lymph node ≥2 cm that is amenable to biopsy.
* Willing and able to give informed consent if the subject is 18 years of age or older, adolescent assent and legal guardian consent for subjects 15-17 years of age, pediatric assent and legal guardian consent for subjects 7-14 years of age, or legal guardian consent for pediatric subjects less than 7 years of age.

Exclusion Criteria:

* Cannot or is not willing to follow study procedures.
* Subjects with a previous history of cancer in the last five years with the exception of subjects with Kaposi sarcoma (i.e. history of Kaposi sarcoma is allowed in the study).

• Michael Painschab, MD, Principal investigator, IUSCCC