Cytokines Expression in Pregnant Women With Primary and Non-primary Cytomegalovirus (CMV) Infection

@#Introduction: Cytomegalovirus (CMV) infection in pregnancy is the commonest cause of congenital infection worldwide. Primary CMV infection in pregnancy carries a higher risk of fetal transmission compared to non-primary infection. This study aims to determine the cytokines expression in pregnant women with primary and non-primary CMV infections in both types of infection. Methods: This prospective cohort study was conducted at Microbiology Laboratory, Universiti Sains Malaysia (USM) from January 2019 until June 2020. Seventy-four pregnant women with abnormal pregnancy outcomes with positive CMV IgG with or without IgM by electrochemiluminescence assay (ECLIA) were subjected to IgG avidity assay by ECLIA method to discriminate primary and non-primary CMV infection. Later, the sera were subjected to magnetic Luminex multiplex enzyme-linked immunosorbent assay for cytokine analysis to determine their concentrations in both primary and non-primary CMV infection. Cytokines and chemokines tested were IL-12, IL-2, IFN- γ, TNF-α, IL-1β, IL-6, IL-10, IFN- γ, TNF-α, MCP-1 (CCL-2), and IP-10 (CXCL-10). Results: Concentrations of IL-1β, IL-6, and MCP-1 (CCL-2) were significantly elevated in pregnant women with primary CMV infection with the p-values of (0.001, 0.035, and 0.002) respectively. The intensity of IFN-γ, IL-12, and IL-2 were higher in primary CMV infection with the p-values of (0.018, 0.004, and 0.007). Conclusion: The pro-inflammatory cytokines were expressed significantly in pregnant women with primary CMV infection together with MCP-1 (CCL2), showing predominant Th1 response. The low level of cytokines in non-primary CMV infection might be due to the latent state of CMV in a host.

Mini Review on Laboratory Practices During Covid-19 Pandemic: From Microbiologists Perspective

@#Laboratory practices in a laboratory have changed worldwide due to the emergence of the COVID-19 pandemic. The changes occur concerning specimen collection, handling, transportation, processing, and disposal. Infection control practices are applied in all aspects, starting from specimen collection until the clinician gets the results. A retrospective review of laboratory practices used in a tertiary teaching hospital laboratory from microbiologists’ perspectives was performed, and the practices were compared with previously published articles.

Diagnostic Use of PCR in Carbapenamase-producing Enterobacteriaceae (CPE): An Improved Method to Overcome the presence of inhibitors for DNA Extraction from Blood Cultures

@#Introduction: Carpanenamase-producing Enterobacteriaceae (CPE) has emerged as a threat to hospitalized patients. Phenotypic test such as Modified hodge test was less sensitive and specific especially to detect blaNDM-1 which is the most predominant genotype in this region. Nucleic acid amplification technology offers improved specificity and sensitivity. Failed amplification due to the presence of inhibitors is a limitation. In this study, we tried to use previous method described by Villumseen et al with some modification using another DNA extraction kit. Methods: Ten mls of sterile whole blood taken from nearly expired blood bag from blood bank was spiked with 200 µl of 0.5mcFarland bacterial suspension from thirty-six confirmed isolates of blaNDM-1 carbapenamase-producing Klebsiella pneumoniae in an aerobic Bactec Plus and incubated until the growth was detected. The blood specimen was subjected to DNA extraction method using Macherey-Nachel, Nucleospin® Blood QuickPure followed with multiplex PCR. Results: Out of the 36 isolates, 12 isolates revealed blaNDM-1 , 9 isolates revealed blaNDM-1 and blaOXA-48, 7 isolates revealed blaNDM-1, blaVIM and blaKPC genotypes that were amplified at cycle threshold of less than 30. Another 8 isolates could not pick up any genotypes possibly due to pipetting error as all the internal control were amplified. Eight true negative gram negative isolates underwent same procedure and none amplified at a cycle threshold less than 30. Conclusion: This modified method was proved to give a high yield of CPE genotypes with the cycle threshold was set at less than or equal to 30 and able to overcome the presence of PCR inhibitors

Cardiac Tamponade: A Rare Complication of Plasmodium Knowlesi Malaria

Abstract@#Plasmodium knowlesi has been discovered as the fifth species causing malaria in humans. It is a major public health problem in South East Asia especially in Borneo. We report a case of pericardial effusion that rapidly progressing to cardiac tamponade, an atypical presentation of P. knowlesi malaria. Our patient had no underlying known medical illness, presented with high grade fever with chills and rigors, epigastric pain, nausea, vomiting and with poor oral intake. Initial bedside cardiac ultrasound showed minimal pericardial effusion. Within a few hours, she became hypotensive, deteriorated rapidly despite fluid resuscitation requiring mechanical ventilation and inotropic support. Bedside cardiac ultrasound showed cardiac tamponade and pericardiocentesis was done. We highlight the importance of having high level of suspicion for this atypical presentation of cardiac tamponade when a patient is hypotensive in P. knowlesi infection. Prompt diagnosis and management may prevent potentially fatal complication.

Diagnostic Use of PCR in Carbapenamase-producing Enterobacteriaceae (CPE): An Improved Method to Overcome the presence of inhibitors for DNA Extraction from Blood Cultures

@#Introduction: Carpanenamase-producing Enterobacteriaceae (CPE) has emerged as a threat to hospitalized patients. Phenotypic test such as Modified hodge test was less sensitive and specific especially to detect blaNDM-1 which is the most predominant genotype in this region. Nucleic acid amplification technology offers improved specificity and sensitivity. Failed amplification due to the presence of inhibitors is a limitation. In this study, we tried to use previous method described by Villumseen et al with some modification using another DNA extraction kit. Methods: Ten mls of sterile whole blood taken from nearly expired blood bag from blood bank was spiked with 200 µl of 0.5mcFarland bacterial suspension from thirty-six confirmed isolates of blaNDM-1 carbapenamase-producing Klebsiella pneumoniae in an aerobic Bactec Plus and incubated until the growth was detected. The blood specimen was subjected to DNA extraction method using Macherey-Nachel, Nucleospin® Blood QuickPure followed with multiplex PCR. Results: Out of the 36 isolates, 12 isolates revealed blaNDM-1 , 9 isolates revealed blaNDM-1 and blaOXA-48, 7 isolates revealed blaNDM-1, blaVIM and blaKPC genotypes that were amplified at cycle threshold of less than 30. Another 8 isolates could not pick up any genotypes possibly due to pipetting error as all the internal control were amplified. Eight true negative gram negative isolates underwent same procedure and none amplified at a cycle threshold less than 30. Conclusion: This modified method was proved to give a high yield of CPE genotypes with the cycle threshold was set at less than or equal to 30 and able to overcome the presence of PCR inhibitors.

Awake fibre Optic Intubation with Dexmedetomidine for Ludwig Angina with Severe Trismus

@#Ludwig angina is a submandibular space cellulitis secondary to oral cavity infection. It is strongly associated with difficult intubation due to limitation in the mouth opening. The presentation of Ludwig angina varies according to the severity of the infection. The extreme presentations include upper airway obstruction and respiratory failure. We present a female teenager with right submandibular abscess as the consequence of Ludwig angina, who was planned for incision and drainage. Successful awake fibre optic intubation was performed as a method of induction due to trismus, deferring the need for tracheostomy.