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Fulltext Identification of Mycobacterium tuberculosis-specific DNA marker using MAMA-PCR

Chin Kai Ling, Jaeyres Jani, Zainal Arifin Mustapha

Malaysian Journal of Medicine and Health Sciences, 2019;15(106):36-36.
MyJurnal

Introduction: Tuberculosis (TB), commonly caused by Mycobacterium tuberculosis (Mtb), is one of the ten leading causes of death worldwide. The gold standard, microbiological culture for detection and differentiation of mycobac-teria are time-consuming and laborious. The use of fast, easy and sensitive nucleic acid amplification tests (NAATs) for diagnosis of TB remains challenging because there is a high degree of homology within Mtb complex (MTBC) members and absence of target genes in the genome of some strains. This study aimed to identify new candidate genetic marker and to design specific primers to detect Mtb using in silico methods. Methods: Using Basic Local Alignment Search Tool (BLAST) program, Mtb H37Rv chromosome reference genome sequence was mapped with other MTBC members and a single nucleotide polymorphism (SNP) at Rv1970 was found to be specific only for Mtb strains. Mismatch amplification mutation assay (MAMA) combine with polymerase chain reaction (PCR) was used as an alternative method to detect the point mutation. MAMA primers targeting the SNP were designed using Primer-BLAST and the PCR assay was optimized via Taguchi method. Results: The assay amplified a 112 bp gene fragment and was able to detect all Mtb strains, but not the other MTBC members and non-tuberculous Mycobacte-ria. The detection limit of the assay was 60 pg/μl. Conclusion: Bioinformatics has provided predictive identification of many new target markers. The designed primers were found to be highly specific at single-gene target resolution for detection of Mtb.
Transcriptome analysis and molecular characterization of novel small RNAs in Mycobacterium tuberculosis Lineage 1

Jumat MI, Chin KL

World J Microbiol Biotechnol, 2024 Jul 25;40(9):279.
PMID: 39048776 DOI: 10.1007/s11274-024-04089-6

Mycobacterium tuberculosis (Mtb), the tuberculosis-causing agent, exhibits diverse genetic lineages, with known links to virulence. While genomic and transcriptomic variations between modern and ancient Mtb lineages have been explored, the role of small non-coding RNA (sRNA) in post-translational gene regulation remains largely uncharted. In this study, Mtb Lineage 1 (L1) Sabahan strains (n = 3) underwent sRNA sequencing, revealing 351 sRNAs, including 23 known sRNAs and 328 novel ones identified using ANNOgesic. Thirteen sRNAs were selected based on the best average cut-off value of 300, with RT-qPCR revealing significant expression differences for sRNA 1 (p = 0.0132) and sRNA 29 (p = 0.0012) between Mtb L1 and other lineages (L2 and L4, n = 3) (p > 0.05). Further characterization using RACE (rapid amplification of cDNA ends), followed by target prediction with TargetRNA3 unveils that sRNA 1 (55 base pairs) targets Rv0506, Rv0697, and Rv3590c, and sRNA 29 (86 base pairs) targets Rv33859c, Rv3345c, Rv0755c, Rv0107c, Rv1817, Rv2950c, Rv1181, Rv3610c, and Rv3296. Functional characterization with Mycobrowser reveals these targets involved in regulating intermediary metabolism and respiration, cell wall and cell processes, lipid metabolism, information pathways, and PE/PPE. In summary, two novel sRNAs, sRNA 1 and sRNA 29, exhibited differential expression between L1 and other lineages, with predicted roles in essential Mtb functions. These findings offer insights into Mtb regulatory mechanisms, holding promise for the development of improved tuberculosis treatment strategies in the future.
Fulltext Selection and validation of emotional videos: Dataset of professional and amateur videos that elicit basic emotions

Chen H, Chin KL, Tan CBY

Data Brief, 2021 Feb;34:106662.
PMID: 33376765 DOI: 10.1016/j.dib.2020.106662

This article describes the process of selecting a collection of professional and amateur videos that elicit five basic emotions (i.e., happiness, fear, disgust, anger, and sadness) and validating these videos in three groups of participants (i.e., Chinese from China, Chinese from Malaysia, and Bumiputera from Malaysia). In the video selection phase, professional videos, which were Western movie trailers, were selected from IMDb (Internet Movie Database) and amateur videos were selected from YouTube. The researchers selected videos that display five basic emotions, identified the time frames with the strongest display of emotion, and rated the emotional intensity of each video on a 5-point Likert scale. After the initial stage of selection, two other researchers performed an emotion recognition task by watching the videos without audio to ensure that the emotions can be elicited without understanding the language. This data was used to refine the final selection of 20 professional videos and 20 amateur videos. In the video validation phase, 30 participants were asked to identify and rate the intensity of emotion felt. This article includes a description of the video selection method, a detailed list of the videos selected, and participants' responses and ratings of emotional intensity for the 40 videos.
DNA markers for tuberculosis diagnosis

Chin KL, Sarmiento ME, Norazmi MN, Acosta A

Tuberculosis (Edinb), 2018 12;113:139-152.
PMID: 30514496 DOI: 10.1016/j.tube.2018.09.008

Tuberculosis (TB), caused by Mycobacterium tuberculosis complex (MTBC), is an infectious disease with more than 10.4 million cases and 1.7 million deaths reported worldwide in 2016. The classical methods for detection and differentiation of mycobacteria are: acid-fast microscopy (Ziehl-Neelsen staining), culture, and biochemical methods. However, the microbial phenotypic characterization is time-consuming and laborious. Thus, fast, easy, and sensitive nucleic acid amplification tests (NAATs) have been developed based on specific DNA markers, which are commercially available for TB diagnosis. Despite these developments, the disease remains uncontrollable. The identification and differentiation among MTBC members with the use of NAATs remains challenging due, among other factors, to the high degree of homology within the members and mutations, which hinders the identification of specific target sequences in the genome with potential impact in the diagnosis and treatment outcomes. In silico methods provide predictive identification of many new target genes/fragments/regions that can specifically be used to identify species/strains, which have not been fully explored. This review focused on DNA markers useful for MTBC detection, species identification and antibiotic resistance determination. The use of DNA targets with new technological approaches will help to develop NAATs applicable to all levels of the health system, mainly in low resource areas, which urgently need customized methods to their specific conditions.
Fulltext Challenges and the Way forward in Diagnosis and Treatment of Tuberculosis Infection

Chin KL, Anibarro L, Sarmiento ME, Acosta A

Trop Med Infect Dis, 2023 Jan 28;8(2).
PMID: 36828505 DOI: 10.3390/tropicalmed8020089

Globally, it is estimated that one-quarter of the world's population is latently infected with Mycobacterium tuberculosis (Mtb), also known as latent tuberculosis infection (LTBI). Recently, this condition has been referred to as tuberculosis infection (TBI), considering the dynamic spectrum of the infection, as 5-10% of the latently infected population will develop active TB (ATB). The chances of TBI development increase due to close contact with index TB patients. The emergence of multidrug-resistant TB (MDR-TB) and the risk of development of latent MDR-TB has further complicated the situation. Detection of TBI is challenging as the infected individual does not present symptoms. Currently, there is no gold standard for TBI diagnosis, and the only screening tests are tuberculin skin test (TST) and interferon gamma release assays (IGRAs). However, these tests have several limitations, including the inability to differentiate between ATB and TBI, false-positive results in BCG-vaccinated individuals (only for TST), false-negative results in children, elderly, and immunocompromised patients, and the inability to predict the progression to ATB, among others. Thus, new host markers and Mtb-specific antigens are being tested to develop new diagnostic methods. Besides screening, TBI therapy is a key intervention for TB control. However, the long-course treatment and associated side effects result in non-adherence to the treatment. Additionally, the latent MDR strains are not susceptible to the current TBI treatments, which add an additional challenge. This review discusses the current situation of TBI, as well as the challenges and efforts involved in its control.
Fulltext Antioxioxidant and antiapoptotic effects of Thymosin β4 in Aβ-induced SH-SY5Y cells via the 5-HTR1A/ERK axis

Zhang GH, Chin KL, Yan SY, Pare R

PLoS One, 2023;18(10):e0287817.
PMID: 37788276 DOI: 10.1371/journal.pone.0287817

Alzheimer's disease (AD) is a common amnestic cognitive impairment characterised by β-amyloid (Aβ) plaques deposit in the brain of the elderly. AD is a yet incurable disease due to its unknown exact pathogenesis and unavailability of effective remedies in clinical application. Thymosin β4 (Tβ4) is a housekeeping protein that plays important role in cell proliferation, migration and differentiation. It has the ability to protect and repair neurons however it is still unclear involvement in AD. Therefore, the aim of this study is to elucidate the role and mechanism of Tβ4 in mediating the improvement of AD. AD-like cell model was constructed in neuroblastoma cell line SH-SY5Y treated with Aβ. Overexpression of Tβ4 were done using lentivirus infection and downregulation through siRNA transfection. We performed western blot and flow cytometry to study the apoptosis and standard kits to measure the oxidative stress-associated biomarkers. There is significant increased in viability and decreased apoptosis in Tβ4 overexpression group compared to control. Furthermore, overexpression of Tβ4 suppressed the expression of pro-apoptotic markers such as Caspase-3, Caspase-8, and Bax meanwhile upregulated the expression of anti-apoptotic gene Bcl-2. Tβ4 alleviated oxidative damage by reducing MDA, LDH and ROS and increasing SOD and GSH-PX in Aβ-treated SH-SY5Y cells. We found that Tβ4 inhibit ERK/p38 MAPK pathway and intensify the expression of 5-HTR1A. Additionally, we showed that upregulation of 5-HTR1A dampened the Tβ4 to activate ERK signalling. In conclusion, our study revealed the neuroprotective role of Tβ4 in AD which may open up new therapeutic applications in AD treatment.
Role of non-coding RNAs in tuberculosis and their potential for clinical applications

Jumat MI, Sarmiento ME, Acosta A, Chin KL

J Appl Microbiol, 2023 Jun 01;134(6).
PMID: 37197901 DOI: 10.1093/jambio/lxad104

Tuberculosis (TB), caused by Mycobacterium tuberculosis (Mtb), remains the leading cause of mortality due to infectious diseases, only surpassed in 2020 by COVID-19. Despite the development in diagnostics, therapeutics, and evaluation of new vaccines for TB, this infectious disease remains uncontrollable due to the emergence of multidrug-resistant (MDR) and extremely drug-resistant (XDR) TB, among other factors. The development in transcriptomics (RNomics) has enabled the study of gene expression in TB. It is considered that non-coding RNAs (ncRNAs) from host [microRNAs (miRNAs)] and Mtb [small RNAs (sRNAs)] are important elements in TB pathogenesis, immune resistance, and susceptibility. Many studies have shown the importance of host miRNAs in regulating immune response against Mtb via in vitro and in vivo mice models. The bacterial sRNAs play a major role in survival, adaptation, and virulence. Here, we review the characterization and function of host and bacteria ncRNAs in TB and their potential use in clinical applications as diagnostic, prognostic, and therapeutic biomarkers.
Fulltext Aptamer-Based Strategies to Address Challenges in COVID-19 Diagnosis and Treatments

Chang ZY, Alhamami FAMS, Chin KL

Interdiscip Perspect Infect Dis, 2023;2023:9224815.
PMID: 37554129 DOI: 10.1155/2023/9224815

Coronavirus disease (COVID-19), a highly contagious and rapidly spreading disease with significant fatality in the elderly population, has swept across the world since 2019. Since its first appearance, the causative agent, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has undergone multiple mutations, with Omicron as the predominant circulating variant of concern at the moment. The gold standard for diagnosis of COVID-19 by real-time polymerase chain reaction (RT-PCR) to detect the virus is laborious and requires well-trained personnel to perform sophisticated procedures. Also, the genetic variants of SARS-CoV-2 that arise regularly could result in false-negative detection. Meanwhile, the current COVID-19 treatments such as conventional medicine, complementary and alternative medicine, passive antibody therapy, and respiratory therapy are associated with adverse effects. Thus, there is an urgent need to discover novel diagnostic and therapeutic approaches against SARS-CoV-2 and its variants. Over the past 30 years, nucleic acid-based aptamers have gained increasing attention and serve as a promising alternative to the antibodies in the diagnostic and therapeutic fields with their uniqueness of being small, nonimmunogenicity, and thermally stable. Aptamer targeting the SARS-CoV-2 structural proteins or the host receptor proteins represent a powerful tool to control COVID-19 infection. In this review, challenges faced by currently available diagnostic and therapeutic tools for COVID-19 are underscored, along with how aptamers can shed a light on the current COVID-19 pandemic, focusing on the critical factors affecting the discovery of high-affinity aptamers and their potential applications to control COVID-19 infection.
Tβ4 ameliorates oxidative damage and apoptosis through ERK/MAPK and 5-HT1A signaling pathway in Aβ insulted SH-SY5Y cells

Zhang GH, Pare RB, Chin KL, Qian YH

Life Sci, 2021 Nov 25.
PMID: 34838849 DOI: 10.1016/j.lfs.2021.120178

AIMS: Alzheimer's disease (AD) is the most common progressive neurodegenerative disorder seriously endangering the physical and mental health of the elderly, while no effective treatments and drugs in clinical practice are available. Thymosin β4 (Tβ4) is a multifunctional polypeptide involved in many physiological and pathological processes including AD. This study aims to understand the function and molecular mechanism of Tβ4 in the development of AD.
MAIN METHODS: Neuroblastoma cell line SH-SY5Y was treated with β-amyloid (Aβ) to induce AD-like pathological changes, which serves as Alzheimer's disease model. Tβ4 was overexpressed in SH-SY5Y cells by lentivirus infection, and downregulated by siRNA transfection. Apoptosis of transfected SH-SY5Y cells after Aβ-treatment was examined by western blot and flow cytometry. Apoptotic proteins and Tβ4-related signaling pathways were also investigated by western blot.
KEY FINDINGS: We found that Tβ4 overexpression increased viability and suppressed apoptosis of Aβ-treated SH-SY5Y cells. Tβ4 ameliorated oxidative damage and suppressed reactive oxygen species production in Aβ-treated SH-SY5Y cells. Consistently, Tβ4 overexpression down-regulated the expression levels of pro-apoptotic markers such as Caspase-3, Caspase-8, and Bax, while up-regulated the expression level of anti-apoptotic gene Bcl-2 in Aβ-stimulated SH-SY5Y cells. Mechanistically, we demonstrated that Tβ4 dampened ERK/p38 MAPK signaling and enhanced 5-HTR1A expression in Aβ-treated SH-SY5Y cells. Moreover, we revealed that Tβ4 inhibited the activation of ERK pathway through up-regulating 5-HTR1A in Aβ-treated SH-SY5Y cells.
SIGNIFICANCE: Taken together, our findings provide evidences to support the neuroprotective role of Tβ4 and might open up new therapeutic applications of Tβ4 in AD treatment.
Fulltext Correlation of mean cycle threshold value of GeneXpert MTB/RIF and time to detection in Mycobacterium growth indicator tubes for Mycobacterium tuberculosis isolates of Kota Kinabalu, Sabah

Nor Fazirah Jamal, Zaw Lin, Zainal Arifin Mustafa, Ahmad Faris Abdullah, Naing Oo Tha, Nor Amalina Emran, et al.

Malaysian Journal of Medicine and Health Sciences, 2019;15(106):38-38.
MyJurnal

Introduction: Tuberculosis (TB) is the ninth leading cause of mortality in the world while it is the most prevalent infection which is ranked abolve HIV/AIDS. In Malaysia, tuberculosis is still a public health problem. Sabah State on Malaysian Borneo had 20-30% of total TB cases of the country. In Sabah, case notification rate of almost 200 per 100,000 population was still present in the last 10 years. Hotspots are defined as TB notification rate more than 100/100.000 in a district or TB notification rate more than 100/100,000 in the squatters’ area. In this study, cycle threshold (ct) values in GeneXpertMTB/RIF (Xpert) were tried to be correlated with growth in Mycobacterim growth indicator tubes (MGIT) by measurement of time to detection (TTD). Methods: Sputum samples from six hotspots of Kota Kinabalu were studied by Xpert as well as MGIT culture after processing of sputum samples. The correlation between Mean ct value of Xpert and TTD in MGIT was analysed by using IBM SPSS Statistic 25 and the statistical test used was Pearson Correlation test. Results: The results of Xpert indicated 35 of 204 sputum samples were pos-itive whereas only one sample was rifampicin resistant. The mean ct values were very low, low and medium in all the hotspots with sputum from one hotspot showed medium ct values predominantly. The sputum from remaining hotspots showed mainly very low and low ct values. MGIT results showed no growth in five samples with two very low, two low and one medium mean ct values. Conclusion: The finding indicated that there were correlations be-tween mean ct values of Xpert and TTD in MGIT with a few exceptions.
Fulltext Brain-Derived Neurotrophic Factor Polymorphism and Aphasia after Stroke

Lee NT, Ahmedy F, Mohamad Hashim N, Yin KN, Chin KL

Behav Neurol, 2021;2021:8887012.
PMID: 34367374 DOI: 10.1155/2021/8887012

Stroke is one of the most deliberating causes of mortality and disability worldwide. Studies have implicated Val66Met polymorphism of the brain-derived neurotrophic factor (BDNF) gene as a genetic factor influencing stroke recovery. Still, the role of BDNF polymorphism in poststroke aphasia is relatively unclear. This review assesses the recent evidence on the association between the BDNF polymorphism and aphasia recovery in poststroke patients. The article highlights BNDF polymorphism characteristics, speech and language interventions delivered, and the influence of BNDF polymorphism on poststroke aphasia recovery. We conducted a literature search through PubMed and Google Scholar with the following terms: "brain derived-neurotrophic factor" and "aphasia" for original articles from January 2000 until June 2020. Out of 69 search results, a detailed selection process produced a total of 3 articles that met the eligibility criteria. All three studies included Val66Met polymorphism as the studied human BDNF gene. One of the studies demonstrated insufficient evidence to conclude that BDNF polymorphism plays a role in poststroke aphasia recovery. The remaining two studies have shown that Met allele genotype (either single or double nucleotides) was associated with poor aphasia recovery, in either acute or chronic stroke. Carriers of the Val66Met polymorphism of BDNF gave a poorer response to aphasia intervention and presented with more severe aphasia.
Fulltext Mitochondrial DNA sequence of the horseshoe crab Tachypleus gigas

Sarmiento ME, Chin KL, Lau NS, Aziah I, Norazmi MN, Acosta A, et al.

Mitochondrial DNA B Resour, 2021 May 23;6(6):1710-1714.
PMID: 34104748 DOI: 10.1080/23802359.2021.1930213

This paper reports on the complete mitochondrial (mt) genome of a horseshoe crab, Tachypleus gigas (T. gigas), in Kuala Kemaman, Terengganu, Malaysia. Whole-genome sequencing of hemocyte DNA was performed with Illumina HiSeq system and the generated reads were de novo assembled with ABySS 2.1.5 and reassembled using mitoZ against Carcinoscorpius rotundicauda and Limulus polyphemus, resulting in a contig of 15 Kb. Phylogenetic analysis of the assembled mt genome suggests that the Tachypleus gigas is closely related to Tachypleus tridentatus than to Carcinoscorpius rotundicauda.
Identification of a Mycobacterium tuberculosis-specific gene marker for diagnosis of tuberculosis using semi-nested melt-MAMA qPCR (lprM-MAMA)

Chin KL, Sarmiento ME, Mustapha ZA, Jani J, Jamal NB, Stanis CS, et al.

Tuberculosis (Edinb), 2020 12;125:102003.
PMID: 33099253 DOI: 10.1016/j.tube.2020.102003

Tuberculosis (TB) is the deadliest of infectious diseases. TB diagnosis, based on sputum microscopy, culture, and nucleic acid amplification tests (NAATs) to identify its main causative agent, Mycobacterium tuberculosis (MTB), remains challenging. The current available NAATs, endorsed by World Health Organization (WHO), can differentiate MTB from some MTB complex (MTBC) members. Using bioinformatics, we identified a single nucleotide polymorphism (SNP) in lprM (Rv1970) gene that differentiate MTB from other MTBC members. A forward mismatch amplification mutation assay (MAMA) primer was designed for the targeted mutation and was used in a semi-nested melt-MAMA qPCR (lprM-MAMA). Using the optimized protocol, lprM-MAMA was positive with all MTB reference and clinical strains, and negative with other MTBC members, non-tuberculous mycobacteria (NTM) and other non-mycobacterial (NM) reference strains. The limit of detection (LOD) of lprM-MAMA was 76.29 fg. Xpert® MTB/RIF (Xpert)-positive sputum samples were also positive by lprM-MAMA, except for samples classified as having "very low" bacterial load by Xpert. Xpert-negative sputum samples were also negative by lprM-MAMA. In conclusion, lprM-MAMA demonstrated to be a useful tool for specific MTB diagnosis. Further evaluation with higher number of reference strains, including NTM and NM; and sputum samples are required to determine its potential for clinical application.
Comparative transcriptome profiling of horseshoe crab Tachypleus gigas hemocytes in response to lipopolysaccharides

Sarmiento ME, Chin KL, Lau NS, Aziah I, Ismail N, Norazmi MN, et al.

Fish Shellfish Immunol, 2021 Oct;117:148-156.
PMID: 34358702 DOI: 10.1016/j.fsi.2021.08.001

Horseshoe crabs (HSCs) are living fossil species of marine arthropods with a long evolutionary history spanning approximately 500 million years. Their survival is helped by their innate immune system that comprises cellular and humoral immune components to protect them against invading pathogens. To help understand the genetic mechanisms involved, the present study utilised the Illumina HiSeq platform to perform transcriptomic analysis of hemocytes from the HSC, Tachypleus gigas, that were challenged with lipopolysaccharides (LPS). The high-throughput sequencing resulted in 352,077,208 and 386,749,136 raw reads corresponding to 282,490,910 and 305,709,830 high-quality mappable reads for the control and LPS-treated hemocyte samples, respectively. Based on the log-fold change of > 0.3 or
Corrigendum to 'Identification of a mycobacterium tuberculosis-specific gene marker for diagnosis of tuberculosis using semi-nested melt-MAMA qPCR (lprM-MAMA)' [Tuberculosis 125 (2020) 102003]

Chin KL, Sarmiento ME, Mustapha ZA, Jani J, Jamal NB, Stanis CS, et al.

Tuberculosis (Edinb), 2021 Sep;130:102121.
PMID: 34474263 DOI: 10.1016/j.tube.2021.102121
Fulltext Detection of Salivary IgA Antibodies Against the HlyE Antigen as a Diagnosis of Typhoid Fever

Chin KL, Redhuan NE, Balaram P, Phua KK, Ong EB

J Clin Diagn Res, 2016 Jun;10(6):DM01-3.
PMID: 27504289 DOI: 10.7860/JCDR/2016/17801.7909

The Salmonella typhi (S. typhi) haemolysin E protein (HlyE) has been shown to be a sensitive and specific antigen for the detection of typhoid fever through the detection of anti-HlyE antibodies in sera. Saliva can also be a useful diagnostic fluid as it also contains antibodies against bacterial pathogens.
Fulltext Salivary Anti-50 kDa Antibodies as a Useful Biomarker for Diagnosis of Typhoid Fever

Redhuan NEM, Chin KL, Adnan AS, Ismail A, Balaram P, Phua KK

J Clin Diagn Res, 2017 Jun;11(6):DC10-DC13.
PMID: 28764158 DOI: 10.7860/JCDR/2017/21928.10055

INTRODUCTION: Typhoid fever remains a scourge of humanity, especially in developing and under-developed countries due to poor sanitation and food hygiene. Diagnostic methods available for detection of this disease are not satisfactory due to a lack of sensitive, specific, rapid and convenient diagnostic test kits available in the market.
AIM: To evaluate the feasibility of a Dot-EIA method for Ig-class specific salivary antibody detection for diagnosis of typhoid fever.
MATERIALS AND METHODS: Paired saliva and serum samples were collected in the year 2010 from patients and normal volunteers in Hospital Universiti Sains Malaysia, Kelantan, Malaysia, which is endemic for typhoid fever. A total of 11 culture-confirmed typhoid fever patients, 43 non-typhoid fever patients and 53 normal human control subjects were evaluated for antibodies against a 50 kDa antigen specific for Salmonella Typhi using Dot-EIA.
RESULTS: Ig class-specific screening of the test samples showed a higher sensitivity for IgA (90.9%) compared to either IgG (72.7%) or IgM (72.7%) antibodies in saliva, but for serum, IgG (90.9%) had a higher degree of sensitivity compared to IgA (36.4%) and IgM (63.6%). Combining all isotypes (IgA, IgG or IgM), serum showed a higher sensitivity (100.0%) compared to saliva (90.9%). Also, the specificity for serum (100.0%) was much higher than saliva (85.4%).
CONCLUSION: Salivary IgA anti-50kDa antibody was found to be more suitable biomarker for routine screening, whereas serum IgG was more suitable for confirmatory test as it has higher specificity. Nevertheless, salivary IgA Dot-EIA is a convenient method for rapid testing, such as for Point-of-Care Diagnostics (POCD) and field epidemiological studies, due to its non-invasive nature and ease of use.
Fulltext Role of Interferons in the Development of Diagnostics, Vaccines, and Therapy for Tuberculosis

Chin KL, Anis FZ, Sarmiento ME, Norazmi MN, Acosta A

J Immunol Res, 2017;2017:5212910.
PMID: 28713838 DOI: 10.1155/2017/5212910

Tuberculosis (TB) is an airborne infection caused by Mycobacterium tuberculosis (Mtb). About one-third of the world's population is latently infected with TB and 5-15% of them will develop active TB in their lifetime. It is estimated that each case of active TB may cause 10-20 new infections. Host immune response to Mtb is influenced by interferon- (IFN-) signaling pathways, particularly by type I and type II interferons (IFNs). The latter that consists of IFN-γ has been associated with the promotion of Th1 immune response which is associated with protection against TB. Although this aspect remains controversial at present due to the lack of established correlates of protection, currently, there are different prophylactic, diagnostic, and immunotherapeutic approaches in which IFNs play an important role. This review summarizes the main aspects related with the biology of IFNs, mainly associated with TB, as well as presents the main applications of these cytokines related to prophylaxis, diagnosis, and immunotherapy of TB.
Fulltext Transcriptomic Signature of Horseshoe Crab Carcinoscorpius rotundicauda Hemocytes' Response to Lipopolysaccharides

Sarmiento ME, Chin KL, Lau NS, Ismail N, Norazmi MN, Acosta A, et al.

Curr Issues Mol Biol, 2022 Nov 25;44(12):5866-5878.
PMID: 36547060 DOI: 10.3390/cimb44120399

Carcinoscorpius rotundicauda (C. rotundicauda) is one of the four species of horseshoe crabs (HSCs). The HSC hemocytes store defense molecules that are released upon encountering invading pathogens. The HSCs rely on this innate immunity to continue its existence as a living fossil for more than 480 million years. To gain insight into the innate mechanisms involved, transcriptomic analysis was performed on isolated C. rotundicauda hemocytes challenged with lipopolysaccharides (LPS), the main components of the outer cell membrane of gram-negative bacteria. RNA-sequencing with Illumina HiSeq platform resulted in 232,628,086 and 245,448,176 raw reads corresponding to 190,326,253 and 201,180,020 high-quality mappable reads from control and LPS-stimulated hemocytes, respectively. Following LPS-stimulation, 79 genes were significantly upregulated and 265 genes were downregulated. The differentially expressed genes (DEGs) were related to multiple immune functional categories and pathways such as those of the cytoskeleton, Toll and Imd, apoptosis, MAP kinase (MAPK), inositol phosphate metabolism, phagosome, leucocyte endothelial migration, and gram-negative bacterial infection, among others. This study provides important information about the mechanisms of response to LPS, which is relevant for the understanding the HSCs' immune response.
Fulltext Immunomodulation resulting of helminth infection could be an opportunity for immunization against tuberculosis and mucosal pathogens

Chin KL, Fonte L, Lim BH, Sarmiento ME, Acosta A

Front Immunol, 2023;14:1091352.
PMID: 37020538 DOI: 10.3389/fimmu.2023.1091352

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