Low-cost conventional PCR techniques enable simultaneous detection of bacterial sexually transmitted infections with enhanced sensitivity and specificity.

PURPOSE: Neisseria gonorrhoeae (NG), Chlamydia trachomatis (CT) and Mycoplasma hominis (MH), the three most common treatable bacterial sexually transmitted infections (STIs) worldwide can lead to many complications if remain untreated. Screening of high-risk population with highly sensitive methods will lead to significant improvement in patient outcomes and will prevent downward transmission. The advantages of Polymerase chain reaction (PCR) based assay are not only high sensitivity and specificity, but also detection of multiple organisms in a single reaction which reduce the result turn-around time. The aim of the present study was to evaluate the feasibility of a multiplex PCR assay method targeting 16S rRNA gene for simultaneous detection of NG, CT and MH infection along with their trend and occurrence among high-risk population in Assam, Northeast India. METHODS: A cross-sectional study was undertaken, where a total of 200 randomly selected patients from high-risk population were included. After validation of singleplex PCR, Multiplex PCR (M-PCR) was performed along with the traditional culture method for NG. RESULTS & CONCLUSION: The overall agreement of M-PCR with singleplex PCR was very high (100%). The occurrence of STI was found to be very high (101/200; 50.5%). Furthermore, co-infection was detected in 10/200; 5%) individuals. Infection was more common among young individuals (p < 0.05) and males out-numbered females (p < 0.05). The most common organism detected was CT (42/200; 21%) followed by NG (41/200; 20.5%) and MH (20/200; 10%). The M-PCR assay workflow is simple, cost effective and can be used in routine diagnostic laboratories with basic molecular facilities.

Capsular typing of Streptococcus pneumoniae isolated from clinical specimens in Gauhati Medical College and hospital, Assam, India.

PURPOSE: Streptococcus pneumoniae is an important human respiratory tract pathogen causing pneumococcal diseases in majority of children and adults. The capsule is a significant virulence factor of Pneumococci which determines the bacterial serotype and is the component used for synthesis of pneumococcal vaccines. This cross-sectional study aimed to isolate Streptococcus pneumoniae from clinical samples and determine the occurrence of its circulating serotypes in Assam, North East India. MATERIALS AND METHODS: A total of 80 clinical samples were collected from June 2019 to May 2020 from patients clinically suspected from pneumococcal infection and also included samples routinely sent to bacteriology laboratory. Isolation and identification of S. pneumoniae was performed using conventional culture and molecular methods. Antibiotic susceptibility patterns were monitored. Capsular serotyping was performed using PCR of cpsA gene followed by DNA sequencing. RESULTS: Majority of the cases suspected of pneumococcal infection belong to the paediatric group aged less than 5 years. Out of 80 samples, 10 (12.50%) were found to be positive by PCR of recP gene. Culture was positive in 80% (8/10) of the total positives. Co-trimoxazole resistance was seen in 33.33% of the isolate from sputum. Serotypes 6A, 6B, 6C and 19F were detected in our region, out of which 6C is a non-vaccine serotype. CONCLUSION: Continued surveillance is needed to monitor trends in non-vaccine serotypes that may emerge as highly associated with antibiotic resistance. Also, the need to continuous monitoring of the antibiotic susceptibility of S. pneumoniae in North eastern parts of India is of outmost importance.

In-house reverse transcriptase polymerase chain reaction for detection of SARS-CoV-2 with increased sensitivity.

As the COVID-19 infection continues to ravage the world, the advent of an efficient as well as the economization of the existing RT-PCR based detection assay essentially can become a blessing in these testing times and significantly help in the management of the pandemic. This study demonstrated an innovative and rapid corroboration of COVID-19 test based on innovative multiplex PCR. An assessment of optimal PCR conditions to simultaneously amplify the SARS-CoV-2 genes E, S and RdRp has been made by fast-conventional and HRM coupled multiplex real-time PCR using the same sets of primers. All variables of practical value were studied by amplifying known target-sequences from ten-fold dilutions of archived positive samples of COVID-19 disease. The multiplexing with newly designed E, S and RdRp primers have shown an efficient amplification of the target region of SARS-CoV-2. A distinct amplification was observed in 37 min using thermal cycler while it took 96 min in HRM coupled real time detection using SYBR green over a wide range of template concentrations. Our findings revealed decent concordance with other commercially available detection kits. This fast HRM coupled multiplex real-time PCR with SYBR green approach offers rapid and sensitive detection of SARS-CoV-2 in a cost-effective manner apart from the added advantage of primer compatibility for use in conventional multiplex PCR. The highly reproducible novel approach can propel extended applicability for developing sustainable commercial product besides providing relief to a resource limited setting.

An appraisal of clinicopathological parameters in Japanese encephalitis and changing epidemiological trends in upper Assam, India.

CONTEXT: Japanese encephalitis (JE), an acute mosquito-borne viral disease, is one of the leading causes of viral encephalitis in the South-East Asian region. JE is endemic in Assam. The morbidity and mortality due to JE is significant with outbreaks every year during the monsoons. AIMS: The aim was to study the clinicopathological profile of JE; to examine their role in predicting disease outcome; and to document the increase in the incidence of JE among the adult population in this region. MATERIALS AND METHODS: Clinically suspected acute encephalitis syndrome (AES) cases admitted in Assam Medical College and Hospital during the period of May 2011 to April 2012 were tested by JE virus specific Immunoglobulin M capture ELISA. STATISTICAL ANALYSIS USED: Data analysis was performed using SPSS version 16.0. RESULTS: Out of 424 AES cases, 194 were JE positive. The occurrence of JE in adults was higher (P < 0.001) than the pediatric age group. The study recorded a high rate of renal dysfunction in JE cases. A single case of JE induced abortion and two cases of JE-neurocysticercosis co-infections were documented. Regression analysis revealed that adult population, unconsciousness, paresis and elevated cerebrospinal fluid protein level were associated with a worse prognosis in JE cases. Mortality in JE positive cases was higher than the JE negative cases (P = 0.001). CONCLUSION: The study attempts to highlight the role played by a combination of clinical and laboratory parameters in assessing the severity and outcome in JE and may help in directing the limited medical resources toward those that need it the most.