Development of an in-house aPPD ELISA for Mycobacterium avium complex (MAC) antibodies detection in zoo primates

Lekko, Yusuf Madaki and Azlan Che Amat, . and Peck, Toung Ooi and Sharina Omar, . and Siti Zubaidah Ramanoon, . and Mazlina Mazlan, . and Jesse, Faez Firdaus Abdullah (2024) Development of an in-house aPPD ELISA for Mycobacterium avium complex (MAC) antibodies detection in zoo primates. Pertanika Journal Tropical Agricultural Science (Malaysia), 47 (2). pp. 549-557. ISSN 1511-3701

In non-human primates (NHPs), Mycobacterium avium complex (MAC) species are the major source of non-tuberculous mycobacteriosis, causing tuberculous-like lesions in lymph nodes and parenchymatous organs in zoo and wildlife animals. Poor species-specific detection by serological diagnosis has negatively impacted the surveillance of MAC on non-human primates. Serum was collected from suspected twelve (n = 12) NHPs with no record of health monitoring, including gibbon (n = 5), capuchins (n = 2), siamang (n = 2), mandrill (n = 1), and orangutan (n = 2). An in-house avian purified protein derivative (aPPD) enzyme-linked immunosorbent assays (ELISA) antibody detection was developed and modified based on the established protocols. The aPPD ELISA for MAC antibodies detection at serum and Protein-G dilutions of 1:200-0.5µg/ml, respectively, detected 3/12 (25%) positive serum. At both serum and Protein-G dilutions of 1:100-0.05 and 1:300-1 µg/ml, the aPPD ELISA detected 12/12 (100%), respectively. The antibody was not detected for an in-house aPPD ELISA with serum and anti-monkey immunoglobulin G (IgG) dilutions at 1:100-0.5 and 1:300-1 µg/ml. However, 2/12 (16%) was detected using serum and anti-monkey IgG dilutions at 1:200-0.05 µg/ml. An in-house aPPD ELISA procedure for MAC antibodies detection in primates, at serum and Protein-G dilutions of 1:100-0.05 and 1:300-1 µg/ml, both have shown sensitivity and specificity of 100%, positive predictive value and negative predictive value of 100%, respectively. The serum and anti-monkey IgG have shown extremely low sensitivity and specificity. In conclusion, the performance of an in-house aPPD ELISA using three different dilutions on serum and conjugates in detecting MAC in a primate has shown that Protein-G horseradish peroxidase, as secondary conjugates were able to detect MAC antibodies.

Full text available from: Official URL: http://www.pertanika.upm.edu.my/resources/files/Pe...

Abstract

In non-human primates (NHPs), Mycobacterium avium complex (MAC) species are the major source of non-tuberculous mycobacteriosis, causing tuberculous-like lesions in lymph nodes and parenchymatous organs in zoo and wildlife animals. Poor species-specific detection by serological diagnosis has negatively impacted the surveillance of MAC on non-human primates. Serum was collected from suspected twelve (n = 12) NHPs with no record of health monitoring, including gibbon (n = 5), capuchins (n = 2), siamang (n = 2), mandrill (n = 1), and orangutan (n = 2). An in-house avian purified protein derivative (aPPD) enzyme-linked immunosorbent assays (ELISA) antibody detection was developed and modified based on the established protocols. The aPPD ELISA for MAC antibodies detection at serum and Protein-G dilutions of 1:200-0.5µg/ml, respectively, detected 3/12 (25%) positive serum. At both serum and Protein-G dilutions of 1:100-0.05 and 1:300-1 µg/ml, the aPPD ELISA detected 12/12 (100%), respectively. The antibody was not detected for an in-house aPPD ELISA with serum and anti-monkey immunoglobulin G (IgG) dilutions at 1:100-0.5 and 1:300-1 µg/ml. However, 2/12 (16%) was detected using serum and anti-monkey IgG dilutions at 1:200-0.05 µg/ml. An in-house aPPD ELISA procedure for MAC antibodies detection in primates, at serum and Protein-G dilutions of 1:100-0.05 and 1:300-1 µg/ml, both have shown sensitivity and specificity of 100%, positive predictive value and negative predictive value of 100%, respectively. The serum and anti-monkey IgG have shown extremely low sensitivity and specificity. In conclusion, the performance of an in-house aPPD ELISA using three different dilutions on serum and conjugates in detecting MAC in a primate has shown that Protein-G horseradish peroxidase, as secondary conjugates were able to detect MAC antibodies.

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Item Type:	Article
AGROVOC Term:	primates
AGROVOC Term:	zoo animals
AGROVOC Term:	antibodies
AGROVOC Term:	zoonoses
AGROVOC Term:	ELISA
AGROVOC Term:	experimentation
AGROVOC Term:	diagnostic techniques
AGROVOC Term:	radiosensitivity
Geographical Term:	Malaysia
Uncontrolled Keywords:	Antibodies, ELISA, Mycobacterium avium complex, non-human primates, Protein-G
Depositing User:	Ms. Azariah Hashim
Date Deposited:	19 Mar 2026 02:29
Last Modified:	19 Mar 2026 02:29
URI:	http://webagris.upm.edu.my/id/eprint/2936

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