## Protocol Section
### Identification Module
**NCT ID:** NCT02497079

**Brief Title:** Diagnostic Accuracy of Polymerase Chain Reaction for Mycobacterium Tuberculosis Using EBUS-TBNA Samples

**Official Title:** Comparison of the Diagnostic Accuracy of Nested and Real-time Polymerase Chain Reaction for Mycobacterium Tuberculosis Using EBUS-TBNA Samples in Patients With Isolated Intrathoracic Lymphadenopathy

#### Organization Study ID Info
**ID:** PNUH-P-1
#### Organization
**Class:** OTHER
**Full Name:** Pusan National University Hospital
### Status Module
#### Completion Date
**Date:** 2018-12

**Type:** ESTIMATED

#### Expanded Access Info
**Last Known Status:** RECRUITING
#### Last Update Post Date
**Date:** 2015-07-20

**Type:** ESTIMATED

**Last Update Submit Date:** 2015-07-16

**Overall Status:** UNKNOWN

#### Primary Completion Date
**Date:** 2018-12

**Type:** ESTIMATED

#### Start Date
**Date:** 2015-07

**Status Verified Date:** 2015-07

#### Study First Post Date
**Date:** 2015-07-14
**Type:** ESTIMATED
**Study First Submit Date:** 2015-07-08
**Study First Submit QC Date:** 2015-07-13
### Sponsor Collaborators Module

#### Lead Sponsor

**Class:** OTHER

**Name:** Pusan National University Hospital

#### Responsible Party

**Investigator Affiliation:** Pusan National University Hospital

**Investigator Full Name:** Jung Seop Eom

**Investigator Title:** Clinical Assistant Professor

**Type:** PRINCIPAL_INVESTIGATOR

### Oversight Module

**Oversight Has DMC:** False

### Description Module

**Brief Summary:** The purpose of this study is to compare the diagnostic efficacy of nested and realtime polymerase chain reaction for Mycobacterium tuberculosis using EBUS-TBNA samples in patients with isolated intrathoracic lymphadenopathy.

**Detailed Description:** Although endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) has been widely used to perform mediastinal lymph node sampling, little information is available on polymerase chain reaction for Mycobacterium tuberculosis (TB-PCR) using EBUS-TBNA samples in patients with isolated intrathoracic lymphadenopathy. In addition, two methods of TB-PCR (nested PCR and realtime PCR) are available for the detection of Mycobacterium tuberculosis and which method is superior for detection of Mycobacterium tuberculosis has not been established in patients with isolated intrathoracic lymphadenopathy.

Thus, this study was design to compare the diagnostic efficacy of nested and realtime TB-PCR using EBUS-TBNA samples in patients with isolated intrathoracic lymphadenopathy in South Korea, where the incidence of tuberculosis is intermediate (97/100,000 per year).

### Conditions Module

**Conditions:**

- Lymphadenopathy

- Tuberculosis

- Sarcoidosis

**Keywords:**

- Bronchoscopy

- Ultrasonography

- Tuberculous lymphadenitis

- Polymerase chain reaction

- Sensitivity and specificity

### Design Module

#### Design Info

**Allocation:** NON_RANDOMIZED

**Intervention Model:** SINGLE_GROUP

##### Masking Info

**Masking:** NONE

**Primary Purpose:** DIAGNOSTIC

#### Enrollment Info

**Count:** 100

**Type:** ESTIMATED

**Phases:**

- NA

**Study Type:** INTERVENTIONAL

### Arms Interventions Module

#### Arm Group 1

**Description:** In all patients, nested polymerase chain reaction for Mycobacterium tuberculosis is performed with formalin-fixed paraffin-embedded specimens obtained by endobronchial ultrasound-guided transbronchial needle aspiration.

**Intervention Names:**

- Device: Nested PCR for formalin-fixed tissues

**Label:** Nested PCR for formalin-fixed tissues

**Type:** EXPERIMENTAL

#### Arm Group 2

**Description:** In all patients, nested polymerase chain reaction for Mycobacterium tuberculosis is performed with specimens in sterile saline obtained by endobronchial ultrasound-guided transbronchial needle aspiration.

**Intervention Names:**

- Device: Nested PCR for fresh tissues

**Label:** Nested PCR for fresh tissues

**Type:** EXPERIMENTAL

#### Arm Group 3

**Description:** In all patients, real-time polymerase chain reaction for Mycobacterium tuberculosis is performed with specimens in sterile saline obtained by endobronchial ultrasound-guided transbronchial needle aspiration.

**Intervention Names:**

- Device: Real-time PCR for fresh tissues

**Label:** Real-time PCR for fresh tissues

**Type:** EXPERIMENTAL

### Interventions

#### Intervention 1

**Arm Group Labels:**

- Nested PCR for formalin-fixed tissues

**Description:** For nested PCR, formalin-fixed paraffin-embedded tissues were incubated in 1 mL of xylene at 60°C for 30 min and then centrifuged for 10 min. Paraffin and the supernatant were removed from the samples after centrifugation. The same procedures were repeated until deparaffinization was complete. After adding 1 mL of alcohol, the samples were centrifuged for 5 min, and the supernatant was removed. The sample was then air-dried as a pellet. DNA was extracted using QIAamp (Qiagen, Valencia, CA, USA). PCR amplifications were performed using Mycobacterium tuberculosis IS6110 primers. The first round using outer primers and the second round using inner primers amplified a 256- and 181-bp fragment, respectively. Finally, the PCR products were visualized in a 2% agarose gel.

**Name:** Nested PCR for formalin-fixed tissues

**Other Names:**

- MTB-PCR kit; Biosewoom, Seoul, South Korea

**Type:** DEVICE

#### Intervention 2

**Arm Group Labels:**

- Nested PCR for fresh tissues

**Description:** For nested PCR with specimens in sterile saline, DNA was extracted using QIAamp (Qiagen, Valencia, CA, USA). PCR amplifications were performed using Mycobacterium tuberculosis IS6110 primers. The first round using outer primers and the second round using inner primers amplified a 256- and 181-bp fragment, respectively. Finally, the PCR products were visualized in a 2% agarose gel.

**Name:** Nested PCR for fresh tissues

**Other Names:**

- MTB-PCR kit; Biosewoom, Seoul, South Korea

**Type:** DEVICE

#### Intervention 3

**Arm Group Labels:**

- Real-time PCR for fresh tissues

**Description:** For real-time PCR, specimens in sterile saline were filtered and 1 mL of phosphate based saline was added. After centrifugation for 3 min, the supernatant was removed. Next, 50 μL of extraction buffer was added to the sample, and the sample was heated at 100°C for 20 min. After centrifugation for 3 min, the supernatant was used in PCR. Real-time PCR was performed using the AdvanSure TB/NTM real-time PCR kit. Three channels were used in the real-time PCR reaction (Mycobacterium tuberculosis complex, mycobacteria, and internal control). Signals for FAM, HEX, and Cy5 were measured in each channel. Mycobacterium tuberculosis was considered present if the cycle threshold of rpoB was less than 35 on each signal and greater than or equivalent to that of IS6110.

**Name:** Real-time PCR for fresh tissues

**Other Names:**

- AdvanSure TB/NTM real-time PCR kit; LG, Seoul, South Korea

**Type:** DEVICE

### Outcomes Module

#### Primary Outcomes

**Description:** In all participants with isolated intrathoracic lymphadenopathy, three modalities of TB-PCR (nested PCR for formalin-fixed paraffin-embedded tissues, nested PCR for fresh tissues and real-time PCR for fresh tissues) are performed using EBUS-TBNA samples. The numbers of participants who reported as positive for each TB-PCR modalities are collected. From these data, sensitivity, specificity and diagnostic accuracy of each TB-PCR modalities to diagnose tuberculous lymphadenitis will be calculated.

**Measure:** Number of participants who detected each polymerase chain reactions for Mycobacterium tuberculosis

**Time Frame:** 6 months

### Eligibility Module

**Eligibility Criteria:** Inclusion Criteria:

* Patients with isolated intrathoracic lymphadenopathy

Exclusion Criteria:

* Suspicion of primary lung cancer on CT or PET scan
* Suspicion of pulmonary tuberculosis on CT or PET scan
* Suspicion of sarcoidosis of lung parenchyma on CT or PET scan

**Minimum Age:** 18 Years

**Sex:** ALL

**Standard Ages:**

- ADULT

- OLDER_ADULT

### Contacts Locations Module

#### Central Contacts

**Contact 1:**

**Email:** ejspulm@gmail.com

**Name:** Jung Seop Eom Eom, Master

**Phone:** 82-01-2081-0859

**Role:** CONTACT

#### Locations

**Location 1:**

**City:** Busan

**Contacts:**

***Contact 1:***

- **Email:** ejspulm@gmail.com

- **Name:** Jung Seop Eom, Master

- **Phone:** +821020810859

- **Role:** CONTACT

***Contact 2:***

- **Email:** mokgamokga@gmail.com

- **Name:** Jeong Ha Mok, Master

- **Phone:** +820185196183

- **Role:** CONTACT

**Country:** Korea, Republic of

**Facility:** Pusan National University Hospital

**Status:** RECRUITING

**Zip:** 602-739

#### Overall Officials

**Official 1:**

**Affiliation:** Pusan National University Hospital

**Name:** Jeong Ha Mok, Master

**Role:** PRINCIPAL_INVESTIGATOR

**Official 2:**

**Affiliation:** Pusan National University Hospital

**Name:** Jung Seop Eom, Master

**Role:** PRINCIPAL_INVESTIGATOR

### References Module

#### References

**Citation:** Navani N, Lawrence DR, Kolvekar S, Hayward M, McAsey D, Kocjan G, Falzon M, Capitanio A, Shaw P, Morris S, Omar RZ, Janes SM; REMEDY Trial Investigators. Endobronchial ultrasound-guided transbronchial needle aspiration prevents mediastinoscopies in the diagnosis of isolated mediastinal lymphadenopathy: a prospective trial. Am J Respir Crit Care Med. 2012 Aug 1;186(3):255-60. doi: 10.1164/rccm.201203-0393OC. Epub 2012 May 31.

**PMID:** 22652031

**Citation:** Navani N, Molyneaux PL, Breen RA, Connell DW, Jepson A, Nankivell M, Brown JM, Morris-Jones S, Ng B, Wickremasinghe M, Lalvani A, Rintoul RC, Santis G, Kon OM, Janes SM. Utility of endobronchial ultrasound-guided transbronchial needle aspiration in patients with tuberculous intrathoracic lymphadenopathy: a multicentre study. Thorax. 2011 Oct;66(10):889-93. doi: 10.1136/thoraxjnl-2011-200063. Epub 2011 Aug 3.

**PMID:** 21813622

**Citation:** Sun J, Teng J, Yang H, Li Z, Zhang J, Zhao H, Garfield DH, Han B. Endobronchial ultrasound-guided transbronchial needle aspiration in diagnosing intrathoracic tuberculosis. Ann Thorac Surg. 2013 Dec;96(6):2021-7. doi: 10.1016/j.athoracsur.2013.07.005. Epub 2013 Sep 12.

**PMID:** 24035300

**Citation:** Statement on sarcoidosis. Joint Statement of the American Thoracic Society (ATS), the European Respiratory Society (ERS) and the World Association of Sarcoidosis and Other Granulomatous Disorders (WASOG) adopted by the ATS Board of Directors and by the ERS Executive Committee, February 1999. Am J Respir Crit Care Med. 1999 Aug;160(2):736-55. doi: 10.1164/ajrccm.160.2.ats4-99. No abstract available.

**PMID:** 10430755

**Citation:** Bezabih M, Mariam DW, Selassie SG. Fine needle aspiration cytology of suspected tuberculous lymphadenitis. Cytopathology. 2002 Oct;13(5):284-90. doi: 10.1046/j.1365-2303.2002.00418.x.

**PMID:** 12421444

## Derived Section

### Condition Browse Module - Ancestors

- ID: D000009164

- Term: Mycobacterium Infections

- ID: D000000193

- Term: Actinomycetales Infections

- ID: D000016908

- Term: Gram-Positive Bacterial Infections

- ID: D000001424

- Term: Bacterial Infections

- ID: D000001423

- Term: Bacterial Infections and Mycoses

- ID: D000007239

- Term: Infections

- ID: D000008232

- Term: Lymphoproliferative Disorders

- ID: D000008206

- Term: Lymphatic Diseases

- ID: D000006968

- Term: Hypersensitivity, Delayed

- ID: D000006967

- Term: Hypersensitivity

- ID: D000007154

- Term: Immune System Diseases

### Condition Browse Module - Browse Branches

- Abbrev: BC15

- Name: Blood and Lymph Conditions

- Abbrev: All

- Name: All Conditions

- Abbrev: BC20

- Name: Immune System Diseases

- Abbrev: BC01

- Name: Infections

- Abbrev: BC23

- Name: Symptoms and General Pathology

### Condition Browse Module - Browse Leaves

- ID: M994

- Name: Lymphadenopathy

- Relevance: HIGH

- As Found: Lymphadenopathy

- ID: M10018

- Name: Hypersensitivity

- Relevance: LOW

- As Found: Unknown

- ID: M12119

- Name: Mycobacterium Infections

- Relevance: LOW

- As Found: Unknown

- ID: M17127

- Name: Tuberculosis

- Relevance: HIGH

- As Found: Tuberculosis

- ID: M15325

- Name: Sarcoidosis

- Relevance: HIGH

- As Found: Sarcoidosis

- ID: M17138

- Name: Tuberculosis, Lymph Node

- Relevance: LOW

- As Found: Unknown

- ID: M11196

- Name: Lymphadenitis

- Relevance: LOW

- As Found: Unknown

- ID: M10283

- Name: Infections

- Relevance: LOW

- As Found: Unknown

- ID: M6368

- Name: Communicable Diseases

- Relevance: LOW

- As Found: Unknown

- ID: M4722

- Name: Bacterial Infections

- Relevance: LOW

- As Found: Unknown

- ID: M19252

- Name: Gram-Positive Bacterial Infections

- Relevance: LOW

- As Found: Unknown

- ID: M12136

- Name: Mycoses

- Relevance: LOW

- As Found: Unknown

- ID: M4721

- Name: Bacterial Infections and Mycoses

- Relevance: LOW

- As Found: Unknown

- ID: M11225

- Name: Lymphoproliferative Disorders

- Relevance: LOW

- As Found: Unknown

- ID: M11203

- Name: Lymphatic Diseases

- Relevance: LOW

- As Found: Unknown

- ID: M10019

- Name: Hypersensitivity, Delayed

- Relevance: LOW

- As Found: Unknown

- ID: M10200

- Name: Immune System Diseases

- Relevance: LOW

- As Found: Unknown

### Condition Browse Module - Meshes

- ID: D000014376

- Term: Tuberculosis

- ID: D000012507

- Term: Sarcoidosis

- ID: D000072281

- Term: Lymphadenopathy

### Intervention Browse Module - Ancestors

- ID: D000004202

- Term: Disinfectants

- ID: D000000890

- Term: Anti-Infective Agents

### Intervention Browse Module - Browse Branches

- Abbrev: Infe

- Name: Anti-Infective Agents

- Abbrev: CNSDep

- Name: Central Nervous System Depressants

- Abbrev: All

- Name: All Drugs and Chemicals

### Intervention Browse Module - Browse Leaves

- ID: M3777

- Name: Ethanol

- Relevance: LOW

- As Found: Unknown

- ID: M8681

- Name: Formaldehyde

- Relevance: HIGH

- As Found: Encephalomyelitis

- ID: M7383

- Name: Disinfectants

- Relevance: LOW

- As Found: Unknown

- ID: M4214

- Name: Anti-Infective Agents

- Relevance: LOW

- As Found: Unknown

### Intervention Browse Module - Meshes

- ID: D000005557

- Term: Formaldehyde

### Misc Info Module

- Version Holder: 2024-05-24
