OBJECTIVETo establish a one-step four multiplex reverse transcription polymerase chain reaction (RT-PCR) method based on Homo-Tag Assisted Non-Dimer System (HAND) system for simultaneous detection of 4 diarrhea viruses of rotavirus, astrovirus, norovirus and sapovirus.

METHODSPrimers were designed according to the conserved genome sequence of the 4 viruses and the homologous tail sequences were added to the 5' end. The multiplex RT-PCR system was constructed by optimizing the PCR parameters such as the concentration of universal tag primer and genome-specific Homo-Tailed primers. The specificity, stability and sensitivity of the method were evaluated systematically.

RESULTSThe 4 multiplex RT-PCR methods based on HAND system was established successfully. Specificity analysis showed no cross reaction between the 4 diarrhea viruses. The sensitivity analysis showed detection limits for rotavirus, astrovirus, norovirus and sapovirus of 48, 1.92, 9.6 and 48 pg per reaction, respectively.

CONCLUSIONThe established HAND system-based multiplex RT-PCR assay allows simple, rapid, specific, sensitive, and stable for detection of the 4 common diarrhea viruses at low costs and is suitable for application in general medical laboratories.

Astroviridae ; genetics ; isolation & purification ; Diarrhea ; virology ; Feces ; virology ; Humans ; Multiplex Polymerase Chain Reaction ; methods ; Norovirus ; genetics ; isolation & purification ; RNA, Viral ; isolation & purification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Rotavirus ; genetics ; isolation & purification ; Sapovirus ; genetics ; isolation & purification

BackgroundCurrently， there has been a gradual increase in the incidence of mental diseases， becoming a problem that has attracted great concern of the general public. Zhuhai is a core city of the Guangdong-Hong Kong-Macao Greater Bay Area， and the local government have attached great importance to the mental health of residents. The systematic study on the mental health has focused largely on pregnant women and medical workers， yet few are on the adult residents. ObjectiveTo investigate the prevalence and its influencing factors of anxiety and depression among adult residents in Zhuhai， so as to provide references for formulating targeted psychological intervention strategies. MethodsFrom September 1 to December 1， 2021， a cohort of 5 600 permanent residents aged 18 years and above in Zhuhai were selected through multistage sampling. All participants were subjected to complete Patients' Health Questionnaire Depression Scale-9 item （PHQ-9）， Generalized Anxiety Disorder Scale-7 item （GAD-7） and National Mental Health Literacy Questionnaire based on Wenjuanxing software. Then Logistic regression was employed to identify the factors associated with anxiety and depression among the residents. ResultsAmong the 5 600 residents， depressive symptoms were detected in 842 （15.04%） residents and anxiety symptoms in 579 （10.34%） residents. Logistic regression analysis yielded that female gender （OR=1.488）， living in Doumen district （OR=1.942）， junior college/undergraduate level （OR=3.602）， postgraduate and above （OR=5.904）， irregular meal pattern （OR=3.320）， comorbid chronic diseases （OR=3.244）， disqualification of self-assessment （OR=3.414） and disqualification of mental health skill （OR=2.131） were risk factors for depression symptoms， whereas age ≥25 years old （of all age groups OR<1）， monthly per capita income >20 000 yuan （OR=0.438） and exercising ≥3 times a week （OR=0.431） were protective factors for depression symptoms. Age of 25~34 years old （OR=2.051）， living in Jinwan district （OR=1.729） or Doumen district （OR=1.901）， junior college/undergraduate level （OR=2.955）， postgraduate and above （OR=6.662）， irregular meal pattern （OR=2.741）， comorbid chronic diseases （OR=3.535）， disqualification of self-assessment （OR=3.325） and disqualification of mental health skill （OR=1.838） were risk factors for anxiety symptoms， while monthly per capita income of 6 001~9 000 yuan （OR=0.665） and exercising ≥3 times a week （OR=0.572） were protective factors for anxiety symptoms. ConclusionThe mental health status of adult residents in Zhuhai is affected by various factors， including gender， age， region of residence， education level， meal pattern， exercise frequency and chronic diseases. ［Funded by 2023 Zhuhai Medical Research Project （number， 2220009000227）］